Inhibitors of histone demethylases

ABSTRACT

Compounds of the form 
                         
In which Q is selected from —CH═NR 12 , —W, —CH 2 NHR 13 , —CH═O and —CH(OR 17 ) 2  capable of modulating the activity of histone demethylases (HDMEs), which are useful for prevention and/or treatment of diseases in which genomic dysregulation is involved in the pathogenesis, such as e.g. cancer and formulations and methods of use of such compounds.

RELATED APPLICATIONS

This application is a U.S. National Phase application of InternationalApplication No. PCT/EP2014/053674, filed Feb. 26, 2014, which claimspriority to and the benefit of U.S. Provisional Application No.61/770,058, filed Feb. 27, 2013; Danish Application No. PA 2013 70113,filed Feb. 27, 2013; U.S. Provisional Application No. 61/770,065, filedFeb. 27, 2013; Danish Application No. PA 2013 70114, filed Feb. 27,2013, U.S. Provisional Application No. 61/770,067, filed Feb. 27, 2013;Danish Application No. PA 2013 70115, filed Feb. 27, 2013; and U.S.Provisional Application No. 61/931,126, filed Jan. 24, 2014; thecontents of each of which are incorporated by reference herein in theirentireties.

INCORPORATION BY REFERENCE OF SEQUENCE LISTING

The contents of the text file named “EPIZ002N01US_SEQ_LIST.txt”, whichwas created on Aug. 27, 2014 and is 2.4 KB in size, are herebyincorporated by reference in their entireties.

FIELD OF THE INVENTION

The present invention relates to compounds capable of modulating theactivity of histone demethylases (HDMEs), which compounds are useful forthe prevention and/or the treatment of diseases in which genomicdysregulation is involved in the pathogenesis, such as e.g. cancer.

BACKGROUND OF THE INVENTION

The DNA of eukaryotic cells is packaged into chromatin by winding of theDNA around histone proteins to form nucleosomes, the basic unit ofchromatin. One of the important functions of chromatin is to determineregions of active and silenced transcription by changing the orderedchromatin structure. Such changes have profound effects on cellularfunction since they affect fundamental processes as differentiation,proliferation and apoptosis, and are often referred collectively to as“epigenetic” since they can lead to heritable changes that do notinvolve changes in gene sequences (Quina, A. S. et al. (2006), Biochem.Pharmacol. 72; 1563-1569)

These highly controlled chromatin changes are mediated by alterationshistone proteins associated with DNA in the nucleosome. Most notably,the N-terminal histone tail of Histone H3 and histone H4 are subject tosuch covalent changes, which include changes in methylation,acetylation, phosphorylation and ubiquitination. The addition or removalof these groups on histones is mediated by specific enzymes, e.g.histone methyl transferases and histone demethylases for methyl groups,histone acetyltransferases and histone deacetylases for acetyl groups,etc. In the event that the activity or expression of these “epigenetic”enzymes is not correctly controlled and regulated it may lead todisease. Cancer, in particular, is an area of high importance inrelation to dysregulated epigenetic enzyme activity due to the role ofepigenetics in cell differentiation, proliferation and apoptosis, butepigenetics may also play a role in other diseases like metabolic,inflammatory, neurodegenerative and cardiovascular diseases. Thereforethe selective modulation of aberrant action of epigenetic enzymes mayhold great promise for the treatment of human disease (Kelly, T. K. etal. (2010), Nat. Biotechnol. 28; 1069-1078, and Cloos, P. a. C. et al.(2008), Genes. Dev. 22; 115-1140).

Methylation and demethylation of lysine residues on the histone H3 tailconstitute important epigenetic marks delineating transcriptionallyactive and inactive chromatin. For example, methylation of lysine 9 onhistone H3 (H3K9) is usually associated with epigenetically silencedchromatin (Fischle, W., et. al. (2003), Curr. Opinion Cell Biol. 15,172-83; Margueron, R., et al. (2005), Curr. Opinion Genet. Dev. 15,163-76) while methylation of lysine 4 on histone 3 is associated withtranscriptionally active chromatin. Similarly, the lysine 27 histone H3(H3K27) mark is repressive in its di- and tri-methylated states whereasthe lysine 36 histone H3 mark is found in association with geneactivation (Barski, A. et al. (2007), Cell, 129, 823-37; Vakoc, C. etal. (2006) Mol. Cell. Biol. 26, 9185-95; Wagner, E. J. & Carpenter, P.B. (2012) Nature Mol. Cell Biol 13, 115-26). There are, however, manyexemptions from these general rules of association between methylationstates of epigenetic marks and the effect they have on transcription.

As documented by studies of the SUV39H1 knockout mouse, loss of thetri-methyl variant of the H3K9 mark results in chromosomal aberrationsand predisposes to cancer (Peters, A. H. et al., Cell 107, 323-37,2001). The JMJD2C protein (KDM4C, GASC1) has been identified as aneraser of the H3K9 mark (a histone demethylase) and may thereforepromote cancer if its expression and activity is not tightly controlled(Cloos, P. et al. (2006), Nature 442, 307-11; Klose, R. J. et al.(2006), Nature 442, 312-16; Liu, G. et al. (2009), Oncogene 28,4491-500). For example, JMJD2C has been shown to induce transformedphenotypes like growth factor independent growth, anchorage independentgrowth and mammosphere formation, if it is overexpressed in cells (Liu,G. et al. (2009), Oncogene 28, 4491-500). These findings are supportedby the overexpression of JMJD2C in a range of human tumours likesquamous cell carcinoma, metastatic lung carcinoma, prostate cancer,breast cancer and several others (Yang, Z. Q. et al. (2000) Cancer Res.60, 4735-39; Yang, Z. Q. et al. (2001) Jpn. J. Cancer Res. 92, 423-28;Hu, N. et al. (2005) Cancer Res. 65, 2542-46; Liu, G. et al. (2009)Oncogene 28, 4491-500; Wissmann, M. et al. (2007) Nat. Cell Biol. 9,347-53), indicating the potential importance of JMJD2C as an oncogene.

The JMJD2A protein (KDM4A, JHDM3A) shows similar properties to JMJD2C.JMJD2A shows high sequence identity to JMJD2C in its JmjC catalyticdomain, is an eraser of the H3K9 mark and has also been shown to beoverexpressed in prostate cancer (Cloos, P. Et al., Nature 442, 307-11,2006). JMJD2A has been shown to interact with the estrogen receptoralpha (ER-alpha) and overexpression of JMJD2A enhancesestrogen-dependent transcription and the down-regulation of JMJD2Areduced transcription of a seminal ER-alpha target gene, cyclin D1(Kawazu et al., (2011) PLoS One 6; Berry et al., (2012) Int J Oncol 41).Additionally, it has been shown that catalytically inactive JMJD2A iscompromised in its ability to stimulate ER-alpha mediated transcription,suggesting that inhibitors of JMJD2A may be beneficial for the treatmentof ER-alpha positive breast tumours (Berry et al., (2012) Int J Oncol41).

Likewise, an eraser of the tri-methyl variant of the H3K4 mark, JARID1B(KDM5B, PLU1) has also been identified as potential oncogene. In cancerJARID1B most likely acts as a repressor of tumour repressor genes viaremoval of the H3K4 tri-methylation leading to decreased transcriptionalactivation in the affected chromatin regions. The oncogenic potential ofJARID1B is demonstrated by its stimulation of proliferation in celllines and further validated by shRNA knockdown studies of JARID1Bexpression showing inhibition of proliferation in MCF7 human breastcancer cells, in SW780 and RT4 bladder cancer cells, in A549 and LC319lung cancer cells and in 4T1 mouse tumour cells in vitro and/or in mousexenograft experiments (Yamane K. et al. (2007), Mol. Cell 25, 801-12;Hayami S. et al. (2010) Mol. Cancer 9, 59; Catchpole S et al. (2011),Int. J. Oncol. 38, 1267-77). Finally, JARID1B is overexpressed inprostate cancer and is associated with malignancy and poor prognosis(Xiang Y. et al. (2007) PNAS 104).

JARID1A (KDM5A, RBP2) is also an eraser of the tri- and di-methylvariant of the H3K4 mark. JARID1A is overexpressed in gastric cancer(Zeng et al., (2010) Gastroenterology 138) and its gene is amplified incervix carcinoma (Hidalgo et al, (2005) BMC Cancer 5). It has beensuggested that JARID1A is fine-tuning progesterone receptor expressioncontrol by estrogens (Stratmann and Haendler (2011) FEBS J 278).Together with JARID1B, JARID1A has been implicated in the maintenance ofa slow-growing population of cancer cells that are required forcontinuous tumor growth and that are resistant to cytotoxic and targetedtherapy (Roesch, et al, (2010) Cell 141; Sharma, et al., (2010) Cell141). JARID1A is required for the tumor initiation and progression inRb+/− and Menl-defective mice (Lin, et al., (2011) PNAS 108). Data fromPasini show that JARID1A binds to Polycomb group protein target geneswhich are involved in regulating important cellular processes such asembryogenesis, cell proliferation, and stem cell self-renewal throughthe transcriptional repression of genes determining cell fate decisions(Pasini et al., (2008) Genes & Dev 22). Additionally, JARID1A were alsoshown to binds the PRC2 complex and being regulator of PRC2 target genes(Pasini et al., (2008) Genes & Dev 22).

Another potential oncogene, an eraser of the di-methyl variant of theH3K36 mark, JHDM1B (KDM2B, FBXL10) has been shown to be highly expressedin human cancers (Tzatsos A et al. (2009), PNAS 106 (8), 2641-6; He, J.et al. (2011), Blood 117 (14), 3869-80). Knock-down of FBXL10 causessenescence in mouse embryonic fibroblasts (MEFs), which can be rescuedby expression of catalytic active (but not catalytic inactive) JHDM1B(Pfau R et al. (2008), PNAS 105(6), 1907-12; He J et al. (2008), NatStruct Mol Biol 15, 1169-75). JHDM1B demethylates H3K36me2 on thetumor-suppressor gene Ink4b (p15^(Ink4b)), and thereby silences theexpression of this senescence-mediating gene in MEFs and in leukemiccells (He, J. et al. (2008), Nat Struct Mol Biol 15, 1169-75; He, J. etal. (2011), Blood 117 (14), 3869-80). The catalytic dependency of JHDM1Bis further shown by He et al. as catalytic activity is required fordevelopment of leukemia in a mouse AML model.

Inhibitors of the histone demethylase class of epigenetic enzymes, andin particular the potential oncogenes JARID1B, JARID1A, JMJD2C, JMJD2A,and JHDM1B, would present a novel approach for intervention in cancersand other proliferative diseases. Being one of the most devastatingdiseases, affecting millions of people worldwide, there remains a highneed for efficacious and specific compounds against cancer.

PCT/EP2013/070457 discloses histone demethylase (HDME) inhibitors oractivity modulators.

Embodiments of the invention provide novel series of compounds capableof modulating the activity of histone demethylases, at least some ofwhich compounds are useful for the prevention and/or the treatment ofdiseases in which genomic disregulation is involved in the pathogenesis,such as e.g. cancer. By way of the invention

The inventors have surprisingly found that novel compounds of Formula(I) as defined herein can be used in the treatment of HDME dependentdiseases by inhibiting HDMEs. Inhibiting HDMEs would provide a novelapproach to the prevention and treatment of cancer and otherproliferative diseases. Accordingly, it is an object of the presentinvention to provide compounds that when administered alone oroptionally in combination with anti-neoplastic compounds, increases theefficacy of the treatment of HDME dependent diseases.

Accordingly, a first aspect of the present invention relates to acompound of the Formula (I)

wherein

Q is selected from —CH═NR¹², —W, —CH₂NHR¹³, —CH═O and —CH(OR¹⁷)₂;

A is selected from —CHR²C(O)—, C₁₋₈alkylene, C₂₋₈ alkenylene, C₂₋₈alkynylene, C₃₋₁₀ cycloalkylene, heterocyclylene, heteroarylene andarylene, which alkylene, alkenylene, alkynylene, cycloalkylene,heterocyclylene, heteroarylene and arylene may optionally be substitutedwith one or more R³; with the proviso that when Q is —CH═O, A is notalkynylene;

Y is selected from —H, —NR⁶R⁷, —OR⁷, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³ and may form a cyclicstructure with R²; with the proviso that when Q is —CH═O, Y is notalkynyl;

R¹ is selected from —H, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl; ormore preferably is selected from —H and C₁₋₄ alkyl; or with -A-Y forms anitrogen containing optionally substituted heterocyclic group where theoptional substitution may be C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, orC₃₋₁₀ cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl;

R² is selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, andC₃₋₁₀ cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl, andmay form a cyclic structure with Y;

each R³ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷,—Z—SO₂R⁷, —Z—SO₂NR⁶R⁷ and —Z—COOR⁷, wherein any heterocyclyl may besubstituted with one or more R⁴, and wherein any heteroaryl and any arylmay be substituted with one or more R⁵;

Z is selected from a single bond, C₁₋₄ alkylene, heterocyclylene andC₃₋₆ cycloalkylene;

each R⁴ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₁₀ cycloalkyl, —N(R¹)₂, carbamoyl,and —OH;

each R⁵ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₆ cycloalkyl, —CN, —F, —Cl, —Br,carbamoyl and —OH;

each of R⁶ and R⁷ is independently selected from C₁₋₈ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl,which alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl andaryl may optionally be substituted with one or more independentlyselected R⁸; or, alternatively, R⁶ and R⁷ may together with the N-atomto which they are attached form an N-heterocyclic ring optionallysubstituted with one or more independently selected R⁸;

each R⁸ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹,—Z—OR⁹, halogen, —CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclics, heteroaryl and arylmay optionally be substituted with one or more selected from C₁₋₄ alkyl,C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₃₋₆ cycloalkyl, —Z-heterocyclyl,—Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹, —Z—OR⁹, halogen,—CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹; wherein any heterocyclylmay be further substituted with one or more R⁴ as defined above, andwherein any heteroaryl and any aryl may be further substituted with oneor more R⁵ as defined above, and

each R⁹ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, and —Z-heteroaryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above;

each of R¹⁰ and R¹¹ is independently selected from —H, C₁₋₆ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl, and aryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above, or, alternatively, R¹⁰ and R¹¹ may together with theN-atom to which they are attached form an optionally 5 to 7 membered,N-heterocyclic ring optionally substituted with one or more R⁴ asdefined above;

with the proviso that Y is not H when A is —CH₂—;

when Q is —CH═NR¹², R¹² is selected from C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-aryl,—Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷, —Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷,—Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷, —Z—SO₂R⁷ and —Z—COOR⁷, which alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and aryl mayoptionally be substituted with one or more R³; when Q is —CH₂NHR¹³, R¹³is selected from hydrogen, —C(O)R⁷, —C(O)C(O)R⁷, —C(O)C(O)OR⁷, C₁₋₈alkyl, C₁₋₄ fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, and—Z-monocyclic-heteroaryl, which alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, and moncyclic-heteroaryl may optionally be substitutedwith one or more independently selected R⁸, or is —CR¹⁴R¹⁵—NR⁶R⁷,—CR¹⁴R¹⁵CN, or —CR¹⁴R¹⁵OR⁷, wherein each of R¹⁴ and R¹⁵ is independentlyselected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl and aryl, and wherein R¹⁴ and R¹⁵together with the intervening carbon atom may designate a C₃₋₁₀cycloalkyl or C₅₋₁₀-cycloalkenyl ring, which alkyl, alkenyl, alkynyl,cycloalkyl (ring), cycloalkenyl ring, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³;

when Q is W, W is selected from an 1,3-diaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³, and optionally containing one or two oxo groups; a1,3-thiaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³ and optionallycontaining one or two oxo groups; an 1,3-oxaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³, and optionally containing one or two oxo groups, whereinin all three instances two R³'s on the same carbon atom may togetherform a spiro group;

R¹⁶ is selected from hydrogen, —C(O)R⁷, —C(O)C(O)R⁷ and —C(O)C(O)OR⁷;

when Q is —CH(OR¹⁷)₂, each R¹⁷ independently is R³, or wherein two R¹⁷substituents together with the intervening —O—CH(−)—O— may form aheterocyclyl optionally substituted with one or more R³ and containingup to two oxo groups;

or an isomer or a mixture of isomers thereof, or a pharmaceuticallyacceptable salt, or solvate or prodrug thereof.

It is considered to be probable that each of the groups Q is convertedin vivo to produce the corresponding acid (Q=—C(O)OH) by processes whichpossibly include or consist of enzymatic processing. Accordingly, manyor all of the compounds of this invention may act in vivo at leastprincipally in the form of corresponding acid derivatives described inPCT/EP2013/070457. It is thought likely that the enzymatic processingtakes place partly or entirely within cells into which the respectivecompound of the invention has penetrated. In view of this, it isprobable that differences in activity in vitro seen in compounds of theinvention that have the same -A-Y substituent but differ in the group Qare due to the influence of the different groups Q on cell penetrationand/or the efficiency of conversion to the acid form within the cell.This tentative conclusion is based on detection of the correspondingacid within cells following administration of certain compoundsaccording to the invention and molecular modelling of the interaction ofthe acids with relevant enzymes.

Accordingly, in an alternative aspect, the invention provides a compoundof the general Formula

wherein R¹, A, and Y are as defined above or below herein and Q is agroup that is converted to —COOH or COO upon administration of saidcompound to a human, provided that Q is not an amide or an ester of sucha —COOH group.

According to a first set of embodiments, the invention provides acompound of the Formula (I)

wherein

Q is selected from —CH═NR¹² and —W;

A is selected from —CHR²C(O)—, C₂₋₈ alkylene, C₂₋₈ alkenylene, C₂₋₈alkynylene, C₃₋₁₀ cycloalkylene, heterocyclylene, heteroarylene andarylene, which alkylene, alkenylene, alkynylene, cycloalkylene,heterocyclylene, heteroarylene and arylene may optionally be substitutedwith one or more R³;

Y is selected from —H, —NR⁶R⁷, —OR⁷, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³;

R¹ is selected from —H and C₁₋₄ alkyl;

R² is selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl;

each R³ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷,—Z—SO₂R⁷, —Z—SO_(2N)R⁶R⁷ and —Z—COOR⁷, wherein any heterocyclyl may besubstituted with one or more R⁴, and wherein any heteroaryl and any arylmay be substituted with one or more R⁵;

Z is selected from a single bond, C₁₋₄ alkylene, heterocyclylene andC₃₋₆ cycloalkylene;

each R⁴ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₁₀ cycloalkyl, —N(R¹)₂, carbamoyl,and —OH;

each R⁵ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₆ cycloalkyl, —CN, —F, —Cl, —Br,carbamoyl and —OH;

each of R⁶ and R⁷ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl,which alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl andaryl may optionally be substituted with one or more independentlyselected R⁸; or, alternatively, R⁶ and R⁷ may together with the N-atomto which they are attached form an N-heterocyclic ring optionallysubstituted with one or more independently selected R⁸;

each R⁸ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹,—Z—OR⁹, halogen, —CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclics, heteroaryl and arylmay optionally be substituted with one or more selected from C₁₋₄ alkyl,C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₃₋₆ cycloalkyl, —Z-heterocyclyl,—Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹, —Z—OR⁹, halogen,—CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹; wherein any heterocyclylmay be further substituted with one or more R⁴ as defined above, andwherein any heteroaryl and any aryl may be further substituted with oneor more R⁵ as defined above, and

each R⁹ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, and —Z-heteroaryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above;

each of R¹⁰ and R¹¹ is independently selected from —H, C₁₋₆ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl, and aryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above, or, alternatively, R¹⁰ and R¹¹ may together with theN-atom to which they are attached form an N-heterocyclic ring optionallysubstituted with one or more R⁴ as defined above;

R¹² is selected from C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀cycloalkyl, —Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷,—Z—C(═O)—NR⁶R⁷, —Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷,—Z—SOR⁷, —Z—SO₂R⁷ and —Z—COOR⁷, which alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, heteroaryl and aryl may optionally besubstituted with one or more R³;

W is selected from an 1,3-diaza-C₅₋₇-cycloalk-2-yl group which isN-substituted with R¹⁶ and optionally further substituted with one ormore R³, and an 1,3-oxaza-C₅₋₇-cycloalk-2-yl group which isN-substituted with R¹⁶ and optionally further substituted with one ormore R³, wherein in both instances two R³'s on the same carbon atom maytogether form a spiro group;

R¹⁶ is selected from hydrogen, —C(O)R⁷, and —C(O)C(O)R⁷;

with the proviso that Y is not H when A is —CH₂—;

or an isomer or a mixture of isomers thereof, or a pharmaceuticallyacceptable salt, solvate or prodrug thereof.

According to a second set of embodiments, the invention provides acompound of the Formula (I)

wherein

Q is —CH₂NHR¹³;

A is selected from —CHR²C(O)—, C₁₋₈ alkylene, C₂₋₈ alkenylene, C₂₋₈alkynylene, C₃₋₁₀ cycloalkylene, heterocyclylene, heteroarylene andarylene, which alkylene, alkenylene, alkynylene, cycloalkylene,heterocyclylene, heteroarylene and arylene may optionally be substitutedwith one or more R³;

Y is selected from —H, —NR⁶R⁷, —OR⁷, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³;

R¹ is selected from —H and C₁₋₄ alkyl;

R² is selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl;

each R³ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷,—Z—SO₂R⁷, —Z—SO_(2N)R⁶R⁷ and —Z—COOR⁷, wherein any heterocyclyl may besubstituted with one or more R⁴, and wherein any heteroaryl and any arylmay be substituted with one or more R⁵;

Z is selected from a single bond, C₁₋₄ alkylene, heterocyclylene andC₃₋₆ cycloalkylene;

each R⁴ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₁₀ cycloalkyl, —N(R¹)₂, carbamoyl,and —OH;

each R⁵ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₆ cycloalkyl, —CN, —F, —Cl, —Br,carbamoyl and —OH;

each of R⁶ and R⁷ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl,which alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl andaryl may optionally be substituted with one or more independentlyselected R⁸; or, alternatively, R⁶ and R⁷ may together with the N-atomto which they are attached form an N-heterocyclic ring optionallysubstituted with one or more independently selected R⁸;

each R⁸ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹,—Z—OR⁹, halogen, —CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclics, heteroaryl and arylmay optionally be substituted with one or more selected from C₁₋₄ alkyl,C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₃₋₆ cycloalkyl, —Z-heterocyclyl,—Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹, —Z—OR⁹, halogen,—CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹; wherein any heterocyclylmay be further substituted with one or more R⁴ as defined above, andwherein any heteroaryl and any aryl may be further substituted with oneor more R⁵ as defined above, and

each R⁹ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, and —Z-heteroaryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above;

each of R¹⁰ and R¹¹ is independently selected from —H, C₁₋₆ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl, and aryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above, or, alternatively, R¹⁰ and R¹¹ may together with theN-atom to which they are attached form an N-heterocyclic ring optionallysubstituted with one or more R⁴ as defined above;

R¹³ is selected from hydrogen, —C(O)R⁷, —C(O)C(O)R⁷, —R⁷,—CR¹⁴R¹⁵—NR⁶R⁷, —CR¹⁴R¹⁵CN, —CR¹⁴R¹⁵OR⁷, wherein each of R¹⁴ and R¹⁵ isindependently selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, and wherein R¹⁴ andR¹⁵ together with the intervening carbon atom may designate a C₃₋₁₀cycloalkyl or C₅₋₁₀-cycloalkenyl ring, which alkyl, alkenyl, alkynyl,cycloalkyl (ring), cycloalkenyl ring, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³;

with the proviso that Y is not H when A is —CH₂—;

or an isomer or a mixture of isomers thereof, or a pharmaceuticallyacceptable salt, solvate or prodrug thereof.

Optionally, Q is required to be different from -A-Y. Optionally, atleast one of Q and -A-Y is not of the form -alkylene-NH-alkylene-aryl,or more specifically is not of the form -alkylene-NH-alkylene-phenyl.For example, one or both of Q and -A-Y may be not of the form—CH₂—NH—(CH₂)_(x)-phenyl, where x is 1-6 and may in particular be 4.

Optionally, Q does not comprise a polycyclic heteroaryl group, and inparticular, Q may not comprise

and optionally may not be

where ‘alkyl’ may be methyl.

According to a third set of embodiments, the invention provides acompound of the Formula (I)

wherein

Q is selected from —CH═O and —CH(OR¹⁷)₂;

A is selected from —CHR²C(O)—, C₁₋₈alkylene, C₂₋₈ alkenylene, C₂₋₈alkynylene, C₃₋₁₀ cycloalkylene, heterocyclylene, heteroarylene andarylene, which alkylene, alkenylene, alkynylene, cycloalkylene,heterocyclylene, heteroarylene and arylene may optionally be substitutedwith one or more R³;

Y is selected from —H, —NR⁶R⁷, —OR⁷, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³;

R¹ is selected from —H and C₁₋₄ alkyl;

R² is selected from —H, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl;

each R³ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷,—Z—SO₂R⁷, —Z—SO_(2N)R⁶R⁷ and —Z—COOR⁷,

wherein any heterocyclyl may be substituted with one or more R⁴, andwherein any heteroaryl and any aryl may be substituted with one or moreR⁵;

Z is selected from a single bond, C₁₋₄ alkylene, heterocyclylene andC₃₋₆ cycloalkylene,

each R⁴ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₁₀ cycloalkyl, —N(R¹)₂, carbamoyl,and —OH;

each R⁵ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₆ cycloalkyl, —CN, —F, —Cl, —Br,carbamoyl and —OH;

each of R⁶ and R⁷ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl,which alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl andaryl may optionally be substituted with one or more independentlyselected R⁸; or, alternatively, R⁶ and R⁷ may together with the N-atomto which they are attached form an N-heterocyclic ring optionallysubstituted with one or more independently selected R⁸;

each R⁸ is independently selected from C₁₋₆alkyl, C₁₋₄ fluoroalkyl, C₁₋₄hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹,—Z—OR⁹, halogen, —CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclics, heteroaryl and arylmay optionally be substituted with one or more selected from C₁₋₄ alkyl,C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₃₋₆ cycloalkyl, —Z-heterocyclyl,—Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹, —Z—OR⁹, halogen,—CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹; wherein any heterocyclylmay be further substituted with one or more R⁴ as defined above, andwherein any heteroaryl and any aryl may be further substituted with oneor more R⁵ as defined above, and

each R⁹ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, and —Z-heteroaryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above;

each of R¹⁰ and R¹¹ is independently selected from —H, C₁₋₆ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl, and aryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above, or, alternatively, R¹⁰ and R¹¹ may together with theN-atom to which they are attached form an N-heterocyclic ring optionallysubstituted with one or more R⁴ as defined above;

each R¹⁷ independently is R³, or wherein two R¹⁷ substituents togetherwith the intervening —O—CH(−)—O— may form a heterocyclyl optionallysubstituted with one or more R³ and containing up to two oxo groups;

with the proviso that Y is not H when A is —CH₂—;

or an isomer or a mixture of isomers thereof, or a pharmaceuticallyacceptable salt, solvate or prodrug thereof.

In this set of embodiments of the invention, optionally -A-Y does notinclude an alkynylene moiety.

Optionally, -A-Y does not comprise a moiety of the formula

or more particularly a moiety of the formula

A in any of the compounds defined by general formula herein may beselected from —CHR²C(O)—, or C₁₋₈ alkylene, or heterocyclylene.

Y in any of the compounds defined by general formula herein may be—NR⁶R⁷.

A in any of the compounds defined by general formula herein may be—CHR²C(O)—.

A in any of the compounds defined by general formula herein may be—CH₂—C(O)—.

Y in any of the compounds defined by general formula herein may be

wherein n is from 1 to 3 and each of R¹⁰ and R¹¹ independently is asdefined in claim 1.

Y in any of the compounds defined by general formula herein may be

for instance

wherein n is from 1 to 3 and each m independently is from 0 to 2.

Y in any of the compounds defined by general formula herein may beselected from heterocyclyl, heteroaryl and aryl, which may be optionallysubstituted with one or more R³.

R¹³ may be H in any of the compounds defined by general formula herein.

Q may be of the formula:

wherein R¹⁸ and R¹⁹ are hydrogen, or together form a1,3-diaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³, and optionallycontaining one or two oxo groups; a 1,3-thiaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³ and optionally containing one or two oxo groups; an1,3-oxaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³, and optionallycontaining one or two oxo groups, wherein in all three instances twoR³'s on the same carbon atom may together form a spiro group.

In some preferred instances, the compound may be one wherein the moiety-A-Y includes 1-3 cyclic moieties selected from monocylic cycloalkyl,monocyclic heterocyclyl, monocylic heteroaryl, dicyclic heteroaryl andmonocyclic aryl.

In preferred aspects of the invention, the compound may be as shown inTable 1 in the Examples section below.

A compound according to the invention may have a molecular weight of130-1,000 g/mol, such as 180-800 g/mol, e.g. 225-600 g/mol or 250-500g/mol.

The invention includes a pharmaceutical composition comprising at leastone compound of Formula (I) as defined in any paragraph hereincontaining such a definition and optionally one or more pharmaceuticallyacceptable excipients, diluents or carriers.

The invention includes such a pharmaceutical composition, whichcomprises one or more further active substances.

The invention includes a compound for use as a medicament which is acompound of the Formula (I)

wherein

Q is selected from —CH═NR¹², —W, —CH₂NHR¹³, —CH═O and —CH(OR¹⁷)₂;

A is selected from —CHR²C(O)—, C₁₋₈alkylene, C₂₋₈ alkenylene, C₂₋₈alkynylene, C₃₋₁₀ cycloalkylene, heterocyclylene, heteroarylene andarylene, which alkylene, alkenylene, alkynylene, cycloalkylene,heterocyclylene, heteroarylene and arylene may optionally be substitutedwith one or more R³; with the proviso that when Q is —CH═O, A is notalkynylene;

Y is selected from —H, —NR⁶R⁷, —OR⁷, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³ and may form a cyclicstructure with R²; with the proviso that when Q is —CH═O, Y is notalkynyl;

R¹ is selected from —H, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl; ormore preferably is selected from —H and C₁₋₄ alkyl; or with -A-Y forms anitrogen containing optionally substituted heterocyclic group where theoptional substitution may be C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, orC₃₋₁₀ cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl;

R² is selected from —H, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, and C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl, andmay form a cyclic structure with Y;

each R³ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷,—Z—SO₂R⁷, —Z—SO₂NR⁶R⁷ and —Z—COOR⁷, wherein any heterocyclyl may besubstituted with one or more R⁴, and wherein any heteroaryl and any arylmay be substituted with one or more R⁵;

Z is selected from a single bond, C₁₋₄ alkylene, heterocyclylene andC₃₋₆ cycloalkylene;

each R⁴ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₁₀ cycloalkyl, —N(R¹)₂, carbamoyl,and —OH;

each R⁵ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₆ cycloalkyl, —CN, —F, —Cl, —Br,carbamoyl and —OH;

each of R⁶ and R⁷ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl,which alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl andaryl may optionally be substituted with one or more independentlyselected R⁸; or, alternatively, R⁶ and R⁷ may together with the N-atomto which they are attached form an N-heterocyclic ring optionallysubstituted with one or more independently selected R⁸;

each R⁸ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹,—Z—OR⁹, halogen, —CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclics, heteroaryl and arylmay optionally be substituted with one or more selected from C₁₋₄ alkyl,C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₃₋₆ cycloalkyl, —Z-heterocyclyl,—Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹, —Z—OR⁹, halogen,—CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹; wherein any heterocyclylmay be further substituted with one or more R⁴ as defined above, andwherein any heteroaryl and any aryl may be further substituted with oneor more R⁵ as defined above, and

each R⁹ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, and —Z-heteroaryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above;

each of R¹⁰ and R¹¹ is independently selected from —H, C₁₋₆ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl, and aryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above, or, alternatively, R¹⁰ and R¹¹ may together with theN-atom to which they are attached form an N-heterocyclic ring optionallysubstituted with one or more R⁴ as defined above;

with the proviso that Y is not H when A is —CH₂—;

when Q is —CH═NR¹², R¹² is selected from C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-aryl,—Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷, —Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷,—Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷, —Z—SO₂R⁷ and —Z—COOR⁷, which alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and aryl mayoptionally be substituted with one or more R³;

when Q is —CH₂NHR¹³, R¹³ is selected from hydrogen, —C(O)R⁷,—C(O)C(O)R⁷, —C(O)C(O)OR⁷, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, —Z-heterocyclyl, and —Z-monocyclic-heteroaryl, which alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, and monocyclic-heteroarylmay optionally be substituted with one or more independently selectedR⁸, or is —CR¹⁴R¹⁵—NR⁶R⁷, —CR¹⁴R¹⁵CN, or —CR¹⁴R¹⁵OR⁷, wherein each ofR¹⁴ and R¹⁵ is independently selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, andwherein R¹⁴ and R¹⁵ together with the intervening carbon atom maydesignate a C₃₋₁₀ cycloalkyl or C₅₋₁₀-cycloalkenyl ring, which alkyl,alkenyl, alkynyl, cycloalkyl (ring), cycloalkenyl ring, heterocyclyl,heteroaryl and aryl may optionally be substituted with one or more R³;

when Q is W, W is selected from an 1,3-diaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³, and optionally containing one or two oxo groups; a1,3-thiaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³ and optionallycontaining one or two oxo groups; an 1,3-oxaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³, and optionally containing one or two oxo groups, whereinin all three instances two R³'s on the same carbon atom may togetherform a spiro group;

R¹⁶ is selected from hydrogen, —C(O)R⁷, —C(O)C(O)R⁷, —C(O)C(O)R⁷;

when Q is —CH(OR¹⁷)₂, each R¹⁷ independently is R³, or wherein two R¹⁷substituents together with the intervening —O—CH(−)—O— may form aheterocyclyl optionally substituted with one or more R³ and containingup to two oxo groups;

or an isomer or a mixture of isomers thereof, or a pharmaceuticallyacceptable salt, or solvate or prodrug thereof.

The invention includes a compound for use in the treatment of a HDMEdependent disease which is of the Formula (I)

wherein

Q is selected from —CH═NR¹², —W, —CH₂NHR¹³, —CH═O and —CH(OR¹⁷)₂;

A is selected from —CHR²C(O)—, C₁₋₈alkylene, C₂₋₈ alkenylene, C₂₋₈alkynylene, C₃₋₁₀ cycloalkylene, heterocyclylene, heteroarylene andarylene, which alkylene, alkenylene, alkynylene, cycloalkylene,heterocyclylene, heteroarylene and arylene may optionally be substitutedwith one or more R³;

Y is selected from —H, —NR⁶R⁷, —OR⁷, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³ and may form a cyclicstructure with R²;

R¹ is selected from —H, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl; ormore preferably is selected from —H and C₁₋₄ alkyl; or with -A-Y forms anitrogen containing optionally substituted heterocyclic group where theoptional substitution may be C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, orC₃₋₁₀ cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl;

R² is selected from —H, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, and C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl, andmay form a cyclic structure with Y;

each R³ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷,—Z—SO₂R⁷, —Z—SO₂NR⁶R⁷ and —Z—COOR⁷, wherein any heterocyclyl may besubstituted with one or more R⁴, and wherein any heteroaryl and any arylmay be substituted with one or more R⁵;

Z is selected from a single bond, C₁₋₄ alkylene, heterocyclylene andC₃₋₆ cycloalkylene;

each R⁴ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₁₀ cycloalkyl, —N(R¹)₂, carbamoyl,and —OH;

each R⁵ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₆ cycloalkyl, —CN, —F, —Cl, —Br,carbamoyl and —OH;

each of R⁶ and R⁷ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl,which alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl andaryl may optionally be substituted with one or more independentlyselected R⁸; or, alternatively, R⁶ and R⁷ may together with the N-atomto which they are attached form an N-heterocyclic ring optionallysubstituted with one or more independently selected R⁸;

each R⁸ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹,—Z—OR⁹, halogen, —CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclics, heteroaryl and arylmay optionally be substituted with one or more selected from C₁₋₄ alkyl,C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₃₋₆ cycloalkyl, —Z-heterocyclyl,—Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹, —Z—OR⁹, halogen,—CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹; wherein any heterocyclylmay be further substituted with one or more R⁴ as defined above, andwherein any heteroaryl and any aryl may be further substituted with oneor more R⁵ as defined above, and

each R⁹ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, and —Z-heteroaryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above;

each of R¹⁰ and R¹¹ is independently selected from —H, C₁₋₆ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl, and aryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above, or, alternatively, R¹⁰ and R¹¹ may together with theN-atom to which they are attached form an N-heterocyclic ring optionallysubstituted with one or more R⁴ as defined above;

with the proviso that Y is not H when A is —CH₂—;

when Q is —CH═NR¹², R¹² is selected from C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-aryl,—Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷, —Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷,—Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷, —Z—SO₂R⁷ and —Z—COOR⁷, which alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and aryl mayoptionally be substituted with one or more R³;

when Q is —CH₂NHR¹³, R¹³ is selected from hydrogen, —C(O)R⁷,—C(O)C(O)R⁷, —C(O)C(O)OR⁷, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈alkenyl, C₂₋₈alkynyl, C₃₋₁₀cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl, which alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and aryl mayoptionally be substituted with one or more independently selected R⁸, oris —CR¹⁴R¹⁵—NR⁶R⁷, —CR¹⁴R¹⁵CN, or —CR¹⁴R¹⁵OR⁷, wherein each of R¹⁴ andR¹⁵ is independently selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, andwherein R¹⁴ and R¹⁵ together with the intervening carbon atom maydesignate a C₃₋₁₀ cycloalkyl or C₅₋₁₀-cycloalkenyl ring, which alkyl,alkenyl, alkynyl, cycloalkyl (ring), cycloalkenyl ring, heterocyclyl,heteroaryl and aryl may optionally be substituted with one or more R³;

when Q is W, W is selected from an 1,3-diaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³, and optionally containing one or two oxo groups; a1,3-thiaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³ and optionallycontaining one or two oxo groups; an 1,3-oxaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³, and optionally containing one or two oxo groups, whereinin all three instances two R³'s on the same carbon atom may togetherform a spiro group;

R¹⁶ is selected from hydrogen, —C(O)R⁷, —C(O)C(O)R⁷, —C(O)C(O)OR⁷;

when Q is —CH(OR¹⁷)₂, each R¹⁷ independently is R³, or wherein two R¹⁷substituents together with the intervening —O—CH(−)—O— may form aheterocyclyl optionally substituted with one or more R³ and containingup to two oxo groups;

or an isomer or a mixture of isomers thereof, or a pharmaceuticallyacceptable salt, or solvate or prodrug thereof.

The invention includes the use of a compound for the preparation of apharmaceutical composition for the treatment of a HDME dependentdisease, which compound is of the Formula (I)

wherein

Q is selected from —CH═NR¹², —W, —CH₂NHR¹³, —CH═O and —CH(OR¹⁷)₂;

A is selected from —CHR²C(O)—, C₁₋₈ alkylene, C₂₋₈ alkenylene, C₂₋₈alkynylene, C₃₋₁₀ cycloalkylene, heterocyclylene, heteroarylene andarylene, which alkylene, alkenylene, alkynylene, cycloalkylene,heterocyclylene, heteroarylene and arylene may optionally be substitutedwith one or more R³;

Y is selected from —H, —NR⁶R⁷, —OR⁷, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³ and may form a cyclicstructure with R²;

R¹ is selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl; ormore preferably is selected from —H and C₁₋₄ alkyl; or with -A-Y forms anitrogen containing optionally substituted heterocyclic group where theoptional substitution may be C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, orC₃₋₁₀ cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl;

R² is selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, andC₃₋₁₀ cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkyl may beoptionally substituted with one or more selected from —OH, aryl, C₁₋₆alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl, andmay form a cyclic structure with Y;

each R³ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷,—Z—SO₂R⁷, —Z—SO₂NR⁶R⁷ and —Z—COOR⁷, wherein any heterocyclyl may besubstituted with one or more R⁴, and wherein any heteroaryl and any arylmay be substituted with one or more R⁵;

Z is selected from a single bond, C₁₋₄ alkylene, heterocyclylene andC₃₋₆ cycloalkylene;

each R⁴ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₁₀ cycloalkyl, —N(R¹)₂, carbamoyl,and —OH;

each R⁵ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₆ cycloalkyl, —CN, —F, —Cl, —Br,carbamoyl and —OH;

each of R⁶ and R⁷ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl,which alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl andaryl may optionally be substituted with one or more independentlyselected R⁸; or, alternatively, R⁶ and R⁷ may together with the N-atomto which they are attached form an N-heterocyclic ring optionallysubstituted with one or more independently selected R⁸;

each R⁸ is independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹,—Z—OR⁹, halogen, —CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclics, heteroaryl and arylmay optionally be substituted with one or more selected from C₁₋₄ alkyl,C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₃₋₆ cycloalkyl, —Z-heterocyclyl,—Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹, —Z—OR⁹, halogen,—CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹; wherein any heterocyclylmay be further substituted with one or more R⁴ as defined above, andwherein any heteroaryl and any aryl may be further substituted with oneor more R⁵ as defined above, and

each R⁹ is independently selected from —H, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, and —Z-heteroaryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above;

each of R¹⁰ and R¹¹ is independently selected from —H, C₁₋₆ alkyl, C₁₋₄fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl, and aryl, wherein any heterocyclylmay be substituted with one or more R⁴ as defined above, and wherein anyheteroaryl and any aryl may be substituted with one or more R⁵ asdefined above, or, alternatively, R¹⁰ and R¹¹ may together with theN-atom to which they are attached form an N-heterocyclic ring optionallysubstituted with one or more R⁴ as defined above;

with the proviso that Y is not H when A is —CH₂—;

when Q is —CH═NR¹², R¹² is selected from C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-aryl,—Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷, —Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷,—Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷, —Z—SO₂R⁷ and —Z—COOR⁷, which alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and aryl mayoptionally be substituted with one or more R³;

When Q is —CH₂NHR¹³, R¹³ is selected from hydrogen, —C(O)R⁷,—C(O)C(O)R⁷, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄ perfluoroalkyl, C₁₋₄hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl and —Z-aryl, which alkyl, alkenyl,alkynyl, cycloalkyl, heterocyclyl, heteroaryl and aryl may optionally besubstituted with one or more independently selected R⁸, or is—CR¹⁴R¹⁵—NR⁶R⁷, —CR¹⁴R¹⁵CN, or —CR¹⁴R¹⁵OR⁷, wherein each of R¹⁴ and R¹⁵is independently selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl, andwherein R¹⁴ and R¹⁵ together with the intervening carbon atom maydesignate a C₃₋₁₀ cycloalkyl or C₅₋₁₀-cycloalkenyl ring, which alkyl,alkenyl, alkynyl, cycloalkyl (ring), cycloalkenyl ring, heterocyclyl,heteroaryl and aryl may optionally be substituted with one or more R³;

when Q is W, W is selected from an 1,3-diaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³, and optionally containing one or two oxo groups; a1,3-thiaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³ and optionallycontaining one or two oxo groups; an 1,3-oxaza-C₅₋₇-cycloalk-2-yl groupwhich is N-substituted with R¹⁶ and optionally further substituted withone or more R³, and optionally containing one or two oxo groups, whereinin all three instances two R³'s on the same carbon atom may togetherform a spiro group;

R¹⁶ is selected from hydrogen, —C(O)R⁷, —C(O)C(O)R⁷, —C(O)C(O)OR⁷;

when Q is —CH(OR¹⁷)₂, each R¹⁷ independently is R³, or wherein two R¹⁷substituents together with the intervening —O—CH(−)—O— may form aheterocyclyl optionally substituted with one or more R³ and containingup to two oxo groups;

or an isomer or a mixture of isomers thereof, or a pharmaceuticallyacceptable salt, or solvate or prodrug thereof.

The invention includes a method of treating a HDME dependent disease ina subject, said method comprises administering to said subject atherapeutically effective amount of at least one compound of Formula (I)as defined in any one of the above paragraphs.

Conditions treatable using compounds or formulations or compositionsaccording to the invention include cancer in the broadest sense,including solid and non-solid tumours. Further details of treatableconditions appear below.

DETAILED DISCLOSURE OF THE INVENTION

The above definitions of the compounds of Formula (I) are referred toherein by the expressions “compounds of Formula (I)” as defined herein,“compound of Formula (I) as defined herein”, or simply “compounds ofFormula (I)”, etc. It should be understood, that such references areintended to encompass not only the above general formula in its statedaspects, but also each and every of the embodiments, etc. discussedabove or in the following. It should also be understood, that unlessstated to the opposite, such references also encompass isomers, mixturesof isomers, isotopic variants, pharmaceutically acceptable salts,solvates and prodrugs of the compounds of Formula (I).

Without being bound by any particular theory, the current results givereasons to believe that each of the values of Q plays an important rolewhen designing compounds capable of modulating the in vivo activity ofhistone demethylases (HDMEs), whilst in each case the group Q istransformed in vivo to —COOH. Additionally, it is believed that thesubstituent combination -A-Y plays a role in establishing affinity forsaid histone demethylases. Furthermore, it is believed that the pyridinenitrogen and the nitrogen atom of Formula (I) also play a role in thebinding of a particular cavity of the histone demethylases where theiron atom lies. It is also believed that the A-Y chain itself, andthrough its substituents, interacts with the area of the demethylaseknown to accommodate the lysine chain of the substrate.

A is typically selected from —CHR²C(O)—, C₁₋₈ alkylene, C₂₋₈ alkenylene,C₂₋₈ alkynylene, C₃₋₁₀ cycloalkylene, heterocyclylene, heteroarylene andarylene.

The alkylene, alkenylene, alkynylene, cycloalkylene, heterocyclylene,heteroarylene and arylene as A may optionally be substituted with one ormore R³ (see further below).

A may be selected from —CHR²C(O)—, C₁₋₈ alkylene, C₃₋₁₀ cycloalkylene,heterocyclylene, heteroarylene and arylene, in particular from—CHR²C(O)—, C₁₋₈ alkylene and heterocyclylene, such as —CHR²C(O)—, orC₁₋₈ alkylene, or heterocyclylene.

Y is typically selected from —H, —NR⁶R⁷, —OR⁷, C₁₋₈ alkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl and aryl. R⁶and R⁷ are exemplified further below.

The alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl andaryl as Y may optionally be substituted with one or more R³ (see furtherbelow);

In some embodiments, Y is —NR⁶R⁷. In one variant type, A is —CHR²C(O)—and Y is —NR⁶R⁷. In another variant type, A is C₁₋₈ alkyl and Y is—NR⁶R⁷. In one scenario within these embodiments and these variants,—NR⁶R⁷ represents an N-heterocyclic ring optionally substituted with oneor more independently selected R⁸, preferably substituted with one totwo independently selected R⁸. In another scenario within theseembodiments and these variants wherein Y is —NR⁶R⁷, one of R⁶ and R⁷represents —H or C₁₋₆ alkyl. In still another scenario within theseembodiment types and these variants wherein Y is —NR⁶R⁷, R⁶ and R⁷ areindependently selected from C₁₋₈alkyl, C₁₋₄ fluoroalkyl, C₁₋₄hydroxyalkyl, C₂₋₈ alkenyl, and C₂₋₈ alkynyl, e.g. such that R⁶ and R⁷are the same. In still another scenario within these embodiment typesand these variants wherein Y is —NR⁶R⁷, one of R⁶ and R⁷ is selectedfrom heterocyclyl, heteroaryl and aryl.

Y may be —H. In such compounds and in others, A may be selected fromC₁₋₈-alkylene, C₂₋₈ alkenylene, C₂₋₈ alkynylene, and C₃₋₁₀cycloalkylene. In such compounds and in others, A may also be selectedfrom heterocyclyl.

Y may be selected from heterocyclyl, heteroaryl and aryl. In suchcompounds and others, A may be selected from C₁₋₈-alkylene, C₂₋₈alkenylene, C₂₋₈ alkynylene, in particular from C₁₋₈ alkylene, such asfrom C₁₋₆ alkylene, in particular from C₁₋₄ alkylene.

R¹ is typically selected from —H and C₁₋₄ alkyl (such as methyl, ethyl,propyl and butyl), in particular from —H and methyl.

R² is typically selected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₁₀ cycloalkyl, which alkyl, alkenyl, alkynyl and cycloalkylmay be optionally substituted with one or more selected from —OH, aryl,C₁₋₆ alkoxy, heteroaryl, aryloxy, heteroaryloxy, F, and C₃₋₆ cycloalkyl.In some embodiments, R² is selected from —H, C₁₋₄ alkyl (such as methyl,ethyl, propyl and butyl) and C₁₋₄ hydroxyalkyl (such as hydroxymethyl,hydroxyethyl, hydroxypropyl and hydroxybutyl), in particular from —H,methyl and hydroxymethyl.

The R³ (possible substituents to some of the meanings of A and Y) istypically independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷,—Z—SO₂R⁷, —Z—SO₂NR⁶R⁷ and —Z—COOR⁷, wherein any heterocyclyl may besubstituted with one or more R⁴, and wherein any heteroaryl and any arylmay be substituted with one or more R⁵.

Z is typically selected from a single bond, C₁₋₄ alkylene,heterocyclylene and C₃₋₆ cycloalkylene. In one embodiment, Z is selectedfrom C₁₋₄ alkylene. In another embodiment, Z is selected from a singlebond. It should be understood that the group Z may appear several timesin Formula (I) and that such Z's are independently selected.

Each R⁴ (possible substituents of heterocyclyl) may be independentlyselected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₁₋₄alkoxy, C₃₋₁₀ cycloalkyl, —N(R¹)₂, carbamoyl, and —OH,

Each R⁵ (possible substituents of heteroaryl and aryl) may beindependently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄hydroxyalkyl, C₁₋₄ alkoxy, C₃₋₆ cycloalkyl, —CN, —F, —Cl, —Br, carbamoyland —OH.

Each of R⁶ and R⁷ (e.g. of the moiety —NR⁶R⁷) may be independentlyselected from —H (in certain aspects), C₁₋₈ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-heteroaryl and —Z-aryl, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more independently selectedR⁸; or, alternatively, R⁶ and R⁷ may together with the N-atom to whichthey are attached form an N-heterocyclic ring optionally substitutedwith one or more independently selected R⁸.

Each R⁸ may be independently selected from C₁₋₆ alkyl, C₁₋₄ fluoroalkyl,C₁₋₄ hydroxyalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹,—Z—OR⁹, halogen, —CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹, whichalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclics, heteroaryl and arylmay optionally be substituted with one or more selected from C₁₋₄ alkyl,C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₃₋₆ cycloalkyl, —Z-heterocyclyl,—Z-heteroaryl, —Z-aryl, —Z—NR¹⁰R¹¹, —Z—C(═O)—NR¹⁰R¹¹, —Z—OR⁹, halogen,—CN, —Z—SR⁹, —Z—SOR⁹, —Z—SO₂R⁹ and —Z—COOR⁹; wherein any heterocyclylmay be further substituted with one or more R⁴ as defined above, andwherein any heteroaryl and any aryl may be further substituted with oneor more R⁵ as defined above.

Each R⁹ may be independently selected from —H, C₁₋₈alkyl, C₁₋₄fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, —Z-heterocyclyl, —Z-aryl, and —Z-heteroaryl, wherein anyheterocyclyl may be substituted with one or more R⁴ as defined above,and wherein any heteroaryl and any aryl may be substituted with one ormore R⁵ as defined above.

Each of R¹⁰ and R¹¹ (of the moiety —NR¹⁰R¹¹) may be independentlyselected from —H, C₁₋₆ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, heteroaryl, andaryl, wherein any heterocyclyl may be substituted with one or more R⁴ asdefined above, and wherein any heteroaryl and any aryl may besubstituted with one or more R⁵ as defined above, or, alternatively, R¹⁰and R¹¹ may together with the N-atom to which they are attached form anN-heterocyclic ring optionally substituted with one or more R⁴ asdefined above.

In some embodiments, Q is —CH═N—R¹². If so, R¹² may be selected fromC₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, —Z—C(═O)—NR⁶R⁷,—Z—NR⁶—C(═O)—R⁷, —Z—C(═O)—R⁷, —Z—OR⁷, halogen, —Z—SR⁷, —Z—SOR⁷, —Z—SO₂R⁷and —Z—COOR⁷, which alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl,heteroaryl and aryl may optionally be substituted with one or more R³.In some embodiments hereof, R¹² is C₁₋₈ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄perfluoroalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₈ cycloalkyl,—Z-heterocyclyl, —Z-aryl, —Z-heteroaryl, —Z—NR⁶R⁷, and —Z—OR⁷, wherein—Z— is a single bond or C₁₋₄ alkylene, which alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, heteroaryl and aryl may optionally besubstituted with one or more R³.

In other embodiments, Q is —W, wherein —W may be an1,3-azo-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³. W may be1,3-diazacyclopent-2-yl (imidazolidin-2-yl), 1,3-diazacyclohex-2-yl(hexahydropyrimidin-2-yl), or 1,3-diazacyclohept-2-yl, for example. TheN-substituent may be selected among those defined for R¹⁶ (see above). Wmay be further substituted with one or more R³, wherein two R³'s on thesame carbon atom may together form a spiro group.

In yet other embodiments, Q is —W, wherein —W may be an1,3-oxaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³. W may be1,3-oxazacyclopent-2-yl, 1,3-oxazacyclohex-2-yl,1,3-oxazacyclohept-2-yl, or 7-oxa-9-azaspiro[4,5]decan-8-yl, forexample. The N-substituent may be selected among those defined for R¹⁶(see above). W may be further substituted with one or more R³, whereintwo R³'s on the same carbon atom may together form a spiro group.

In some embodiments of the above, W may be further substituted with oneor more R³, but is typically not further substituted.

R¹⁶ may be selected from hydrogen, —C(O)R⁷, —C(O)C(O)R⁷, and—C(O)C(O)OR⁷, in particular from hydrogen and —C(O)R⁷.

In some embodiments Q is —CH₂NHR¹³, and R¹³ may be selected fromhydrogen, —C(O)R⁷, —C(O)C(O)R⁷, —R⁷ (in some aspects), —CR¹⁴R¹⁵—NR⁶R⁷,—CR¹⁴R¹⁵CN, —CR¹⁴R¹⁵OR⁷, wherein each of R¹⁴ and R¹⁵ is independentlyselected from —H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, heteroaryl and aryl, and wherein R¹⁴ and R¹⁵together with the intervening carbon atom may designate a C₃₋₁₀cycloalkyl or C₅₋₁₀-cycloalkenyl ring, which alkyl, alkenyl, alkynyl,cycloalkyl (ring), cycloalkenyl ring, heterocyclyl, heteroaryl and arylmay optionally be substituted with one or more R³. In some aspects,rather than —R⁷, R¹³ may be C₁₋₈ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄perfluoroalkyl, C₁₋₄ hydroxyalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀cycloalkyl, —Z-heterocyclyl, and —Z-monocyclic-heteroaryl, which alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, and heteroaryl mayoptionally be substituted with one or more independently selected R⁸.

In some embodiments Q is —CH(OR¹⁷)₂ and each R¹⁷ independently may beR³, or the two R¹⁷ substituents together with the intervening—O—CH(−)—O— may form a heterocyclyl optionally substituted with one ormore R³.

It is to be understood that in the Formula (I), Y is not H when A is—CH₂—. Generally speaking, it is believed to be advantageous if themoiety -A-Y has a certain “size” with respect to the number of atom(disregarding hydrogen atoms) and/or the molecular weight. Also alimited flexibility of the moiety -A-Y appears to play a certain role.

Hence, it is believed that the moiety -A-Y should preferably consist ofat the most 40 heavy atoms, such as at the most 30 heavy atoms, or atthe most 25 heavy atoms, or at the most 20 heavy atoms. Preferably, themoiety -A-Y will consist of at least 3, or at least 4, or at least 8 orat least 10 heavy atoms. In some embodiments, the moiety -A-Y preferablyconsists of 3-40 heavy atoms, such as 4-30 heavy atoms, or 4-25 heavyatoms, or 4-20, or 8-30, or 8-20, or 8-15 heavy atoms. By the term“heavy atom” is meant all atoms in the moiety except the hydrogenatom(s).

Moreover, it is believed that the compounds of Formula (I) shouldpreferably have a molecular weight of at least 130, or at least 150, orat least 180, or at least 250, but not more than 1000, or not more than800, or not more than 500, or not more than 400 and may be within anyrange constructable from these preferred upper and lower limits, such as130-1,000 g/mol, or 150-1,000 g/mol, such as 180-800 g/mol, e.g. 225-600g/mol or 250-500 g/mol, or 250 to 400.

In some embodiments, and in order to introduce a limited flexibility ofthe moiety -A-Y, the moiety includes 1-4 rings, i.e. rings derived fromcycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl and/or aryl. In somevariant, the moiety -A-Y includes 1-3 cyclic moieties selected frommonocylic cycloalkyl, monocyclic heterocyclyl, monocylic heteroaryl,dicyclic heteroaryl and monocyclic aryl.

Small substituents such as alkyls groups or hydroxyl on alkyl chainsalso reduce flexibility and favor certain conformations.

It may be preferable that if -A-Y does not include a ring, it includesat least one, for instance from 1 to 3, branches, each of whichindependently may be of from one heavy atom to six heavy atoms, forinstance from one to three heavy atoms, or from one to two heavy atoms.It is preferred that -A-Y should contain at least one hetero-atom,preferably at least one nitrogen atom or at least one oxygen.

DEFINITIONS

The term “alkyl” as used herein refers to a saturated, straight orbranched hydrocarbon chain. The hydrocarbon chain preferably containsfrom one to 8 carbon atoms (C₁₋₈-alkyl), more preferred from one to sixcarbon atoms (C₁₋₆-alkyl), in particular from one to four carbon atoms(C₁₋₄-alkyl), including methyl, ethyl, propyl, isopropyl, butyl,isobutyl, secondary butyl, tertiary butyl, pentyl, isopentyl, neopentyl,tertiary pentyl, hexyl, isohexyl, heptyl and octyl. In a preferredembodiment “alkyl” represents a C₁₋₄-alkyl group, which may inparticular include methyl, ethyl, propyl, isopropyl, butyl, isobutyl,secondary butyl, and tertiary butyl. Correspondingly, the term“alkylene” means the corresponding biradical (-alkyl-).

The term “cycloalkyl” as used herein refers to a cyclic alkyl group,preferably containing from three to ten carbon atoms (C₃₋₁₀-cycloalkyl),such as from three to eight carbon atoms (C₃₋₈-cycloalkyl), preferablyfrom three to six carbon atoms (C₃₋₆-cycloalkyl), including cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.Furthermore, the term “cycloalkyl” as used herein may also includepolycyclic groups such as for example bicyclo[2.2.2]octyl,bicyclo[2.2.1]heptanyl, decalinyl and adamantyl. Correspondingly, theterm “cycloalkylene” means the corresponding biradical (-cycloalkyl-).

The term “alkenyl” as used herein refers to a straight or branchedhydrocarbon chain or cyclic hydrocarbons containing one or more doublebonds, including di-enes, tri-enes and poly-enes. Typically, the alkenylgroup comprises from two to eight carbon atoms (C₂₋₈-alkenyl), such asfrom two to six carbon atoms (C₂₋₆-alkenyl), in particular from two tofour carbon atoms (C₂₋₄-alkenyl), including at least one double bond.Examples of alkenyl groups include ethenyl; 1- or 2-propenyl; 1-, 2- or3-butenyl, or 1,3-but-dienyl; 1-, 2-, 3-, 4- or 5-hexenyl, or1,3-hex-dienyl, or 1,3,5-hex-trienyl; 1-, 2-, 3-, 4-, 5-, 6-, or7-octenyl, or 1,3-octadienyl, or 1,3,5-octatrienyl, or1,3,5,7-octatetraenyl, or cyclohexenyl. Correspondingly, the term“alkenylene” means the corresponding biradical (-alkenyl-).

The term “alkynyl” as used herein refers to a straight or branchedhydrocarbon chain containing one or more triple bonds, includingdi-ynes, tri-ynes and poly-ynes. Typically, the alkynyl group comprisesof from two to eight carbon atoms (C₂₋₈-alkynyl), such as from two tosix carbon atoms (C₂₋₆-alkynyl), in particular from two to four carbonatoms (C₂₋₄-alkynyl), including at least one triple bond. Examples ofpreferred alkynyl groups include ethynyl; 1- or 2-propynyl; 1-, 2- or3-butynyl, or 1,3-but-diynyl; 1-, 2-, 3-, 4- or 5-hexynyl, or1,3-hex-diynyl, or 1,3,5-hex-triynyl; 1-, 2-, 3-, 4-, 5-, 6-, or7-octynyl, or 1,3-oct-diynyl, or 1,3,5-oct-triynyl, or1,3,5,7-oct-tetraynyl. Correspondingly, the term “alkynylene” means thecorresponding biradical (-alkynyl-).

The terms “halo” and “halogen” as used herein refer to fluoro, chloro,bromo or iodo. Thus a trihalomethyl group represents e.g. atrifluoromethyl group, or a trichloromethyl group. Preferably, the terms“halo” and “halogen” designate fluoro or chloro.

The term “fluoroalkyl” as used herein refers to an alkyl group asdefined herein which is substituted one or more times with one or morefluorohalo, preferably perfluorated. The term “perfluoroalkyl” as usedherein refers to an alkyl group as defined herein wherein all hydrogenatoms are replaced by fluoro atoms. Preferred fluoroalkyl groups includetrifluoromethyl, pentafluoroethyl, etc.

The term “alkoxy” as used herein refers to an “alkyl-O—” group, whereinalkyl is as defined above.

The term “hydroxyalkyl” as used herein refers to an alkyl group (asdefined hereinabove), which alkyl group is substituted one or more timeswith hydroxy. Examples of hydroxyalkyl groups include HO—CH₂—,HO—CH₂—CH₂— and CH₃—CH(OH)—.

The term “oxy” as used herein refers to an “—O—” group.

The term “oxo” as used herein refers to an “═O” group.

The term “amine” as used herein refers to primary (R—NH₂, R≠H),secondary (R₂—NH, R₂≠H) and tertiary (R₃—N, R≠H) amines. A substitutedamine is intended to mean an amine where at least one of the hydrogenatoms has been replaced by the substituent.

The term “carbamoyl” as used herein refers to a “H₂N(C═O)—” group.

The term “aryl”, as used herein, unless otherwise indicated, includescarbocyclic aromatic ring systems derived from an aromatic hydrocarbonby removal of a hydrogen atom. Aryl furthermore includes bi-, tri- andpolycyclic ring systems. Examples of preferred aryl moieties includephenyl, naphthyl, indenyl, indanyl, fluorenyl, biphenyl, indenyl,naphthyl, anthracenyl, phenanthrenyl, pentalenyl, azulenyl, andbiphenylenyl. Preferred “aryl” is phenyl, naphthyl or indanyl, inparticular phenyl, unless otherwise stated. Any aryl used may beoptionally substituted. Correspondingly, the term “arylene” means thecorresponding biradical (-aryl-).

The term “heteroaryl”, as used herein, refers to aromatic groupscontaining one or more heteroatoms selected from O, S, and N, preferablyfrom one to four heteroatoms, and more preferably from one to threeheteroatoms. Heteroaryl furthermore includes bi-, tri- and polycyclicgroups, wherein at least one ring of the group is aromatic, and at leastone of the rings contains a heteroatom selected from O, S, and N.Heteroaryl also include ring systems substituted with one or more oxomoieties. Examples of preferred heteroaryl moieties includeN-hydroxytetrazolyl, N-hydroxytriazolyl, N-hydroxyimidazolyl, furanyl,triazolyl, pyranyl, thiadiazinyl, benzothiophenyl,dihydro-benzo[b]thiophenyl, xanthenyl, isoindanyl, acridinyl,benzisoxazolyl, quinolinyl, isoquinolinyl, phteridinyl, azepinyl,diazepinyl, imidazolyl, thiazolyl, carbazolyl, pyridinyl, pyridazinyl,pyrimidinyl, pyrazolyl, pyrazinyl, tetrazolyl, furyl, thienyl,isoxazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl,benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl,triazinyl, isoindolyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl,benzofurazanyl, benzothiophenyl, benzotriazolyl, benzothiazolyl,benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl,dihydroquinolyl, tetrahydroquinolyl, dihydroisoquinolyl,tetrahydroisoquinolyl, benzofuryl, furopyridinyl, pyrolopyrimidinyl,azaindolyl, pyrazolinyl, and pyrazolidinyl. Non-limiting examples ofpartially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl,1,4-dihydronaphthyl, and 1-octalin. Correspondingly, the term“heteroarylene” means the corresponding biradical (-heteroaryl-).

The term “heterocyclyl” as used herein, refers to cyclic non-aromaticgroups containing one or more heteroatoms selected from O, S, and N,preferably from one to four heteroatoms, and more preferably from one tothree heteroatoms. Heterocyclyl furthermore includes bi-, tri- andpolycyclic non-aromatic groups, and at least one of the rings contains aheteroatom selected from O, S, and N. Heterocyclyl also include ringsystems substituted with one or more oxo moieties. Examples ofheterocyclic groups are oxetane, pyrrolidinyl, pyrrolyl, 3H-pyrrolyl,oxolanyl, furanyl, thiolanyl, thiophenyl, pyrazolyl, pyrazolidinyl,imidazolyl, imidazolidinyl, 3H-pyrazolyl, 1,2-oxazolyl, 1,3-oxazolyl,1,2-thiazolyl, 1,3-thiazolyl, 1,2,5-oxadiazolyl, piperidinyl, pyridinyl,oxanyl, 2-H-pyranyl, 4-H-pyranyl, thianyl, 2H-thiopyranyl, pyridazinyl,1,2-diazinanyl, pyrimidinyl, 1,3-diazinanyl, pyrazinyl, piperazinyl,1,4-dioxinyl, 1,4-dioxanyl, 1,3-diazinanyl, 1,4-oxazinyl, morpholinyl,thiomorpholinyl, 1,4-oxathianyl, benzofuranyl, isobenzofuranyl,indazolyl, benzimidazolyl, quinolinyl, isoquinolinyl, chromayl,isochromanyl, 4H-chromenyl, 1H-isochromenyl, cinnolinyl, quinazolinyl,quinoxalinyl, phthalazinyl, purinyl, naphthyridinyl, pteridinyl,indolizinyl, 1H-pyrrolizinyl, 4H-quinolizinyl andaza-8-bicyclo[3.2.1]octane. Correspondingly, the term “heterocyclylene”means the corresponding biradical (-heterocyclyl-).

The term “N-heterocyclic ring” as used herein, refers to a heterocyclylor a heteroaryl as defined hereinabove having at least one nitrogenatom, and being bound via a nitrogen atom. Examples of suchN-heterocyclic rings are pyrrolidinyl, pyrrolyl, 3H-pyrrolyl, pyrazolyl,pyrazolidinyl, imidazolyl, imidazolidinyl, 3H-pyrazolyl, 1,2-oxazolyl,1,2-thiazolyl, 1,3-thiazolyl, piperidinyl, pyridinyl, pyridazinyl,pyrazinyl, piperazinyl, morpholinyl, pyridinyl, pyridazinyl,pyrimidinyl, pyrazolyl, pyrazinyl, tetrazolyl, etc.

Isomers

The compounds of Formula (I) may exist as geometric isomers (i.e.cis-trans isomers), optical isomers or stereoisomers, such asdiastereomers, as well as tautomers. Accordingly, it should beunderstood that the definition of compounds of Formula (I) includes eachand every individual isomers corresponding to the structural formula:Formula (I), including cis-trans isomers, stereoisomers and tautomers,as well as racemic mixtures of these and pharmaceutically acceptablesalts thereof. Hence, the definition of compounds of Formula (I) is alsointended to encompass all R- and S-isomers of a chemical structure inany ratio, e.g. with enrichment (i.e. enantiomeric excess ordiastereomeric excess) of one of the possible isomers and correspondingsmaller ratios of other isomers.

Diastereoisomers, i.e. non-superimposable stereochemical isomers, can beseparated by conventional means such as chromatography, distillation,crystallization or sublimation. The optical isomers can be obtained byresolution of the racemic mixtures according to conventional processes,for example by formation of diastereoisomeric salts by treatment with anoptically active acid or base. Examples of appropriate acids include,without limitation, tartaric, diacetyltartaric, dibenzoyltartaric,ditoluoyltartaric and camphorsulfonic acid. The mixture of diastereomerscan be separated by crystallization followed by liberation of theoptically active bases from these salts. An alternative process forseparation of optical isomers includes the use of a chiralchromatography column optimally chosen to maximize the separation of theenantiomers. Still another available method involves synthesis ofcovalent diastereoisomeric molecules by reacting compounds of Formula(I) with an optically pure acid in an activated form or an opticallypure isocyanate. The synthesized diastereoisomers can be separated byconventional means such as chromatography, distillation, crystallizationor sublimation, and then hydrolyzed to obtain the enantiomerically purecompound. The optically active compounds of Formula (I) can likewise beobtained by utilizing optically active starting materials and/or byutilizing a chiral catalyst. These isomers may be in the form of a freeacid, a free base, an ester or a salt. Examples of chiral separationtechniques are given in Chiral Separation Techniques, A PracticalApproach, 2^(nd) ed. by G. Subramanian, Wiley-VCH, 2001.

Pharmaceutically Acceptable Salts

The compound of Formula (I) may be provided in any form suitable for theintended administration, in particular including pharmaceuticallyacceptable salts, solvates and prodrugs of the compound of Formula (I).

Pharmaceutically acceptable salts refer to salts of the compounds ofFormula (I), which are considered to be acceptable for clinical and/orveterinary use. Typical pharmaceutically acceptable salts include thosesalts prepared by reaction of the compounds of Formula (I) a mineral ororganic acid or an organic or inorganic base. Such salts are known asacid addition salts and base addition salts, respectively. It will berecognized that the particular counter-ion or multiple counter-ionsforming a part of any salt is not of a critical nature, so long as thesalt as a whole is pharmaceutically acceptable and as long as thecounter-ion does not contribute undesired qualities to the salt as awhole. These salts may be prepared by methods known to the skilledperson. Pharmaceutically acceptable salts are, e.g., those described anddiscussed in Remington's Pharmaceutical Sciences, 17. Ed. Alfonso R.Gennaro (Ed.), Mack Publishing Company, Easton, Pa., U.S.A., 1985 andmore recent editions and in Encyclopedia of Pharmaceutical Technology.

Examples of pharmaceutically acceptable addition salts include acidaddition salts formed with inorganic acids e.g. hydrochloric,hydrobromic, sulfuric, nitric, hydroiodic, metaphosphoric, or phosphoricacid; and organic acids e.g. succinic, maleic, acetic, fumaric, citric,tartaric, benzoic, trifluoroacetic, malic, lactic, formic, propionic,glycolic, gluconic, camphorsulfuric, isothionic, mucic, gentisic,isonicotinic, saccharic, glucuronic, furoic, glutamic, ascorbic,anthranilic, salicylic, phenylacetic, mandelic, embonic (pamoic),ethanesulfonic, pantothenic, stearic, sulfinilic, alginic andgalacturonic acid; and arylsulfonic, for example benzenesulfonic,p-toluenesulfonic, oxalic, methanesulfonic or naphthalenesulfonic acid;and base addition salts formed with alkali metals and alkaline earthmetals and organic bases such as N,N-dibenzylethylenediamine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine(N-methylglucamine), lysine and procaine; and internally formed salts.

Solvates

The compound of Formula (I) may be provided in dissoluble orindissoluble forms together with a pharmaceutically acceptable solventsuch as water, ethanol, and the like. Dissoluble forms may also includehydrated forms such as the mono-hydrate, the dihydrate, the hemihydrate,the trihydrate, the tetrahydrate, and the like.

Isotopic Variations

Elemental symbols and element names are used herein to include isotopesof the named elements. In particular one, some, or all hydrogens may bedeuterium. Radioactive isotopes may be used, for instance to facilitatetracing the fate of the compounds or their metabolic products afteradministration.

Prodrugs

The compound of Formula (I) may be provided as a prodrug. The term“prodrug” used herein is intended to mean a compound which—upon exposureto certain physiological conditions—will liberate the compound ofFormula (I) which then will be able to exhibit the desired biologicalaction. A typical example is a labile carbamate of an amine and afurther example would be a trialkylsilyl ether of an alcohol or atrialkylsilyl ester of an acid, each optionally being trimethylsilyl.

Inhibitory Effect

The inventors have surprisingly found that compounds of Formula (I) asdefined herein have an inhibitory effect on the activity of one or moreHDMEs. In this respect said one or more HDMEs may be any HDME, howeverpreferably the one or more HDMEs are selected from the JmjC (Jumonji)family, more preferably said one or more HDME(s) are HDME of the humanJmjC family and even more preferably are HDME belonging to the KDM6,KDM5, KDM4 or KDM2 families. The present invention also relates to acompound of Formula (I) as defined herein in a method for inhibitingHDMEs. The method includes contacting a cell with a compound of Formula(I). In a related embodiment, the method further provides that thecompound is present in an amount effective to produce a concentrationsufficient to inhibit the demethylation of a histone in the cell.

Thus, preferably in an assay for demethylation of a histone substrate bysaid HDME, then preferred compounds of Formula (I) are compounds capableof reducing or preferably inhibiting said demethylation by said HDME.Said histone substrate may be any histone, but preferably is histone H3or a fragment thereof, even more preferred: a fragment comprising K4,K9, K27, or K36 of H3. Preferably, said inhibition is determined as theIC₅₀ of said compound of Formula (I) in respect of the saiddemethylation assay.

Preferred compounds of Formula (I) which have an IC₅₀ at or below 1 μM,more preferably less than 300 nM, for example less than 100 nM, such asless than 50 nM in respect of demethylation of any of said histonesubstrates by any of said HDME. Thus very preferred compounds of Formula(I) which have an IC₅₀ at or below 1 μM, more preferably less than 500nM, for example less than 100 nM, such as less than 50 nM in respect ofdemethylation of histone H3 methylated at least on one lysine.

In a preferred embodiment IC₅₀ is determined as described in Example 2herein below. Thus, particularly preferred are compounds of Formula (I)which have an IC₅₀ at or below 1 μM, more preferably less than 500 nM,for example less than 100 nM, such as less than 50 nM when said IC₅₀ isdetermined as described in and one of the Examples herein below.

Particularly preferred compounds of Formula (I) are compounds that leadto a decreased tumour size and/or decreased number of metastases whentested in a xenograft model (Morton and Houghton, Nature Protocols, 2(2) 247-250, 2007).

Pharmaceutical Compositions

In one aspect of this invention, there is provided a pharmaceuticalcomposition comprising at, as an active ingredient, at least onecompound of Formula (I) as defined herein and optionally one or morepharmaceutically acceptable excipients, diluents and/or carriers. Thecompounds of Formula (I) may be administered alone or in combinationwith pharmaceutically acceptable carriers, diluents or excipients, ineither single or multiple doses. Suitable pharmaceutically acceptablecarriers, diluents and excipients include inert solid diluents orfillers, sterile aqueous solutions and various organic solvents.

The pharmaceutical compositions may be formulated with pharmaceuticallyacceptable carriers or diluents as well as any other known adjuvants andexcipients in accordance with conventional techniques such as thosedisclosed in Remington: The Science and Practice of Pharmacy, 21stEdition, 2000, Lippincott Williams & Wilkins.

The pharmaceutical compositions formed by combining a compound ofFormula (I) as defined herein with pharmaceutically acceptable carriers,diluents or excipients can be readily administered in a variety ofdosage forms such as tablets, powders, lozenges, syrups, suppositories,injectable solutions and the like. In powders, the carrier is a finelydivided solid such as talc or starch which is in a mixture with thefinely divided active component. In tablets, the active component ismixed with the carrier having the necessary binding properties insuitable proportions and compacted in the shape and size desired.

The pharmaceutical compositions may be specifically prepared foradministration by any suitable route such as the oral and parenteral(including subcutaneous, intramuscular, intrathecal, intravenous andintradermal) route. It will be appreciated that the preferred route willdepend on the general condition and age of the subject to be treated,the nature of the condition to be treated and the active ingredientchosen.

Pharmaceutical compositions for oral administration include solid dosageforms such as capsules, tablets, dragees, pills, lozenges, powders andgranules. Where appropriate, they can be prepared with coatings such asenteric coatings or they can be prepared so as to provide controlledrelease of the active ingredient such as sustained or prolonged releaseaccording to methods well known in the art.

For oral administration in the form of a tablet or capsule, a compoundof Formula (I) as defined herein may suitably be combined with an oral,non-toxic, pharmaceutically acceptable carrier such as ethanol,glycerol, water or the like. Furthermore, suitable binders, lubricants,disintegrating agents, flavouring agents and colourants may be added tothe mixture, as appropriate. Suitable binders include, e.g., lactose,glucose, starch, gelatin, acacia gum, tragacanth gum, sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes or the like.Lubricants include, e.g., sodium oleate, sodium stearate, magnesiumstearate, sodium benzoate, sodium acetate, sodium chloride or the like.Disintegrating agents include, e.g., starch, methyl cellulose, agar,bentonite, xanthan gum, sodium starch glycolate, crospovidone,croscarmellose sodium or the like. Additional excipients for capsulesinclude macrogols or lipids.

For the preparation of solid compositions such as tablets, the activecompound of Formula (I) is mixed with one or more excipients, such asthe ones described above, and other pharmaceutical diluents such aswater to make a solid pre-formulation composition containing ahomogenous mixture of a compound of Formula (I). The term “homogenous”is understood to mean that the compound of Formula (I) is dispersedevenly throughout the composition so that the composition may readily besubdivided into equally effective unit dosage forms such as tablets orcapsules.

Liquid compositions for either oral or parenteral administration of thecompound of Formula (I) include, e.g., aqueous solutions, syrups,elixirs, aqueous or oil suspensions and emulsion with edible oils suchas cottonseed oil, sesame oil, coconut oil or peanut oil. Suitabledispersing or suspending agents for aqueous suspensions includesynthetic or natural gums such as tragacanth, alginate, acacia, dextran,sodium carboxymethylcellulose, gelatin, methylcellulose orpolyvinylpyrolidone.

Pharmaceutical compositions for parenteral administration includesterile aqueous and non-aqueous injectable solutions, dispersions,suspensions or emulsions as well as sterile powders to be reconstitutedin sterile injectable solutions or dispersions prior to use. Forparenteral administration, solutions containing a compound of Formula(I) in sesame or peanut oil, aqueous propylene glycol, or in sterileaqueous solution may be employed. Such aqueous solutions should besuitably buffered if necessary and the liquid diluent first renderedisotonic with sufficient saline or glucose. These particular aqueoussolutions are especially suitable for intravenous, intramuscular,subcutaneous and intraperitoneal administration. The oily solutions aresuitable for intra-articular, intra-muscular and subcutaneous injectionpurposes.

The preparation of all these solutions under sterile conditions isreadily accomplished by standard pharmaceutical techniques well known tothose skilled in the art.

Depot injectable compositions are also contemplated as being within thescope of the present invention.

In addition to the aforementioned ingredients, the compositions of acompound of Formula (I) may include one or more additional ingredientssuch as diluents, buffers, flavouring agents, colourant, surface activeagents, thickeners, preservatives, e.g. methyl hydroxybenzoate(including anti-oxidants), emulsifying agents and the like.

A suitable dosage of the compound of Formula (I) will depend on the ageand condition of the patient, the severity of the disease to be treatedand other factors well known to the practicing physician. The compoundmay be administered for example either orally, parenterally or topicallyaccording to different dosing schedules, e.g. daily or with intervals,such as weekly intervals. In general a single dose will be in the rangefrom 0.01 to 100 mg/kg body weight, preferably from about 0.05 to 75mg/kg body weight, more preferably between 0.1 to 50 mg/kg body weight,and most preferably between 0.1 to 25 mg/kg body weight. The compoundmay be administered as a bolus (i.e. the entire daily dose isadministered at once) or in divided doses two or more times a day.Variations based on the aforementioned dosage ranges may be made by aphysician of ordinary skill taking into account known considerationssuch as weight, age, and condition of the person being treated, theseverity of the affliction, and the particular route of administration.

The compounds of Formula (I) may also be prepared in a pharmaceuticalcomposition comprising one or more further active substances alone, orin combination with pharmaceutically acceptable carriers, diluents, orexcipients in either single or multiple doses. The suitablepharmaceutically acceptable carriers, diluents and excipients are asdescribed herein above, and the one or more further active substancesmay be any active substances, or preferably an active substance asdescribed in the section “combination treatment” herein below.

Clinical Conditions and Other Uses of Compounds

The compounds according to Formula (I) as defined herein are useful fortreatment of a HDME dependent disease, disorder or condition. Thetreatment may include administering to a mammal, preferably a human,more preferably a human suffering from a HDME dependent disease, atherapeutically effective amount of a compound according to Formula (I)as defined herein.

Said HDME may be any HDME, however preferably the HDME of the presentmethod is selected from the JmjC (Jumonji) family, as described in Clooset. al., Genes & Development 22, 1115-1140, 2008, which is incorporatedherein by reference in its entirety. More preferably said HDME is a HDMEof the human JmjC family.

The present invention also relates to a compound of Formula (I) asdefined herein for use in the treatment of a HDME dependent disease,such as for the treatment of cancer.

By the term “HDME dependent disease” is meant any disease characterizedby elevated HDME expression and/or activity in at least in someinstances of the disease, or a disease which is ameliorated by loweringthe activity of HDMEs. Thus, the disease to be treated with theinhibitors of HDME, i.e. compounds of Formula (I), may be aproliferative or hyperproliferative disease, which includes benign ormalignant tumors, for example a proliferative or hyperproliferativedisease selected from the group consisting of a carcinoma of the brain,kidney, liver, adrenal gland, bladder, breast, stomach (for examplegastric tumors), ovaries, esophagus, colon, rectum, prostate, pancreas,lung, vagina, thyroid, sarcoma, glioblastomas, multiple myeloma orgastrointestinal cancer, for example, colon carcinoma or colorectaladenoma, or a tumor of the neck and head, an epidermalhyperproliferation, for example, psoriasis, prostate hyperplasia, aneoplasia, including a neoplasia of epithelial character, includingmammary carcinoma, and a leukemia.

In one embodiment, compounds of Formula (I) as defined herein are usefulin the treatment of one or more cancers. The term “cancer” refers to anycancer caused by the proliferation of neoplastic cells, such as solidtumors, neoplasms, carcinomas, sarcomas, leukemias, lymphomas and thelike. In particular, cancers that may be treated by the compounds,compositions and methods of the invention include, but are not limitedto: Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma,liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung:bronchogenic carcinoma, (squamous cell, undifferentiated small cell,undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar)carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatoushamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cellcarcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach(carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma,insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), smallbowel (adenocarcinoma, lymphoma, carcinoid tumors, Karposi's sarcoma,leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel(adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor,nephroblastoma, lymphoma, leukemia), bladder and urethra (squamous cellcarcinoma, transitional cell carcinoma, adenocarcinoma), prostate(adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonalcarcinoma, teratocarcfnoma, choriocarcinoma, sarcoma, interstitial cellcarcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver:hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma,angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenicsarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma,chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cellsarcoma), multiple myeloma, malignant giant cell tumor chordoma,osteochronfroma (osteocartilaginous exostoses), benign chondroma,chondroblastoma, chondromyxofibroma, osteoid osteoma and giant celltumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma,osteitis deformans), meninges (meningioma, meningiosarcorna,gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma,germinoma [pinealoma], glioblastoma multiform, oligodendroglioma,schwannoma, retinoblastoma, congenital tumors), spinal cord(neurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus(endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervicaldysplasia), ovaries (ovarian carcinoma, serous cystadenocarcinoma,mucinous cystadenocarcinoma, unclassified carcinoma, granulosa-thecalcell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignantteratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma,adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma,squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma),fallopian tubes (carcinoma); Hematologic: blood (acute myeloid leukemia,chronic myeloid leukemia, acute lymphoblastic leukemia, chroniclymphocytic leukemia, myeloproliferative diseases, multiple myeloma,myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma(malignant lymphoma); Skin: malignant melanoma, basal cell carcinoma,squamous cell carcinoma, Karposi's sarcoma, moles dysplastic nevi,lipoma, angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands:neuroblastoma.

In one embodiment, the compounds of Formula (I) as defined herein areuseful in the treatment of one or more cancers selected from the groupconsisting of: leukemias including acute leukemias and chronic leukemiassuch as acute lymphocytic leukemia (ALL), Acute myeloid leukemia (AML),chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML)and Hairy Cell Leukemia; lymphomas such as cutaneous T-cell lymphomas(CTCL), noncutaneous peripheral T-cell lymphomas, lymphomas associatedwith human T-cell lymphotrophic virus (HTLV) such as adult T-cellleukemia/Iymphoma (ATLL), Hodgkin's disease and non-Hodgkin's lymphomas,large-cell lymphomas, diffuse large B-cell lymphoma (DLBCL); Burkitt'slymphoma; mesothelioma, primary central nervous system (CNS) lymphoma;multiple myeloma; childhood solid tumors such as brain tumors,neuroblastoma, retinoblastoma, Wilm's tumor, bone tumors, andsoft-tissue sarcomas, common solid tumors of adults such as head andneck cancers (e.g., oral, laryngeal and esophageal), genito urinarycancers (e.g., prostate, bladder, renal, uterine, ovarian, testicular,rectal and colon), lung cancer, breast cancer, pancreatic cancer,melanoma and other skin cancers, stomach cancer, brain tumors, livercancer and thyroid cancer.

In another very preferred embodiment, the compound of Formula (I) asdefined herein are useful for the treatment of squamous cell carcinomas.Preferably said squamous cell carcinomas are cancers of the carcinomatype of squamous epithelium that may occur in many different organs,including the skin, lips, mouth, esophagus, urinary bladder, prostate,lungs, vagina, and cervix; brain cancer, that is neuroblastoma,glioblastoma and other malignant and benign brain tumors; breast cancer,pancreatic cancer, and multiple myeloma.

In yet another embodiment, the compounds of Formula (I) as definedherein are useful for treatment of brain cancer, tumors of adults suchas head and neck cancers (e.g., oral, laryngeal and esophageal), genitourinary cancers (e.g., prostate, bladder, renal, uterine, ovarian,testicular, rectal and colon), and breast cancer.

Other cancer forms for which the compounds of Formula (I) are useful astreatment can be found in Stedman's Medical Dictionary (LippincottWilliams & Wilkins, 28^(th) Ed., 2005), which is incorporated herein byreference in its entirety.

In still another related embodiment, the disease to be treated bycompounds of Formula (I) as defined herein is selected from persistentproliferative or hyperproliferative conditions such as angiogenesis,such as psoriasis; Kaposi's sarcoma; restenosis, e.g., stent-inducedrestenosis; endometriosis; Hodgkin's disease; leukemia; hemangioma;angiofibroma; eye diseases, such as neovascular glaucoma; renaldiseases, such as glomerulonephritis; malignant nephrosclerosis;thrombotic microangiopathic syndromes; transplant rejections andglomerulopathy; fibrotic diseases, such as cirrhosis of the liver;mesangial cell-proliferative diseases; injuries of the nerve tissue; andinhibiting the re-occlusion of vessels after balloon catheter treatment,for use in vascular prosthetics or after inserting mechanical devicesfor holding vessels open, such as, e.g., stents, as immune-suppressants,as an aid in scar-free wound healing, and treating age spots and contactdermatitis.

The compounds of Formula (I) are suitable as active agents inpharmaceutical compositions that are efficacious particularly fortreating cellular proliferative or hyperproliferative ailments and/orailments associated with dysregulated gene expression. Suchpharmaceutical compositions have a therapeutically effective amount ofthe compound of Formula (I) along with other pharmaceutically acceptableexcipients, carriers, and diluents and. The phrase, “therapeuticallyeffective amount” as used herein indicates an amount necessary toadminister to a host, or to a cell, tissue, or organ of a host, toachieve a therapeutic effect, such as an ameliorating or alternatively acurative effect, for example an anti-tumor effect, e.g. reduction of orpreferably inhibition of proliferation of malignant cancer cells, benigntumor cells or other proliferative cells, or of any other HDME dependentdisease.

Another aspect of the invention is a pharmaceutical compositioncomprising a therapeutically effective amount of at least one compoundof Formula (I) as defined herein, or a pharmaceutically acceptable salt,solvate or prodrug thereof, in combination with at least one furtheranti-neoplastic compound, and a pharmaceutically acceptable excipient,carrier or diluent.

Method of Treatment

In a further aspect the present invention relates to a method oftreating a diseases in a subject, said method comprises administering tosaid subject a therapeutically effective amount of at least one compoundof Formula (I) as defined herein. The disease may be any disease ordisorder as mentioned herein, such as for example mentioned in thesection “HDME dependent diseases”, and the compound may be administeredalone or in a pharmaceutical composition, such as for example mentionedin the section “Pharmaceutical compositions”.

Hence, the invention also relates to a compound of Formula (I) asdefined herein for use as a medicament.

The term “treating” and “treatment”, as used herein, unless otherwiseindicated, refers to reversing, alleviating, inhibiting the process of,or preventing the disease, disorder or condition to which such termapplies, or one or more symptoms of such disease, disorder or conditionand includes the administration of a compound of Formula (I) to preventthe onset of the symptoms or the complications, or alleviating thesymptoms or the complications, or eliminating the disease, condition, ordisorder. Preferably treatment is curative or ameliorating.

In a preferred embodiment of this aspect of the invention the method isa method of treating a HDME dependent disease in a subject, said methodcomprises administering to said subject a therapeutically effectiveamount of a compound of Formula (I) as defined herein to a subject inneed of such treatment. The HDME dependent disease may be any HDMEdependent disease as described herein above. Preferably the HDMEdependent disease is squamous cell carcinomas or any other of the cancerconditions mentioned above.

Hence, the invention also relates to a compound of Formula (I) asdefined herein for use in the treatment of a HDME dependent disease,such as for the treatment of cancer.

Further, the invention relates to the use of a compound of Formula (I)as defined herein for the preparation of a pharmaceutical compositionfor the treatment of a HDME dependent disease.

In one embodiment of the method of treatment of a HDME dependentdisease, the compound of Formula (I) as defined herein is administeredin combination with one or more further active substances. The activesubstances may be any active substances, and preferably an activesubstance as described herein above in the section “combinationtreatment”. More preferably the one or more additional active substancesare selected from the group consisting of anti-proliferative oranti-neoplastic agents.

Combination Treatment

A compound of Formula (I) may also be used to advantage in combinationwith one or more other anti-proliferative or anti-neoplastic agents.Such anti-proliferative agents include, but are not limited to otherHDME inhibitors, proteasome inhibitors, including bortezomib (Valcade)and Carfilzomib, aromatase inhibitors; antiestrogens; topoisomerase Iinhibitors; topoisomerase II inhibitors; microtubule active agents;alkylating agents; histone deacetylase inhibitors; compounds whichinduce cell differentiation processes; cyclooxygenase inhibitors; MMPinhibitors; mTOR inhibitors; antineoplastic antimetabolites; platincompounds; compounds targeting/decreasing a protein tyrosine or serineor threonine kinase activity; compounds targeting/decreasing a lipidkinase activity; compounds targeting/decreasing a carbohydrate kinaseactivity and further anti-angiogenic compounds; compounds which target,decrease or inhibit the activity of a protein or lipid phosphatase;gonadorelin agonists; anti-androgens; angiostatic steroids; methionineaminopeptidase inhibitors; bisphosphonates; biological responsemodifiers; antiproliferative antibodies; heparanase inhibitors;inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasomeinhibitors; agents used in the treatment of hematologic malignancies;compounds which target, decrease or inhibit the activity of Flt-3; Hsp90inhibitors; temozolomide (TEMOD AL(R)); leucovorin; immune stimulatingagents, such as BCG, IL-2 or IFN-α, antibodies such as anti-CTLA-4monoclonal antibody ipilimumab (Yervoy), rituximab or herceptin andcancer vaccines; inhibitors/modulators of mitochondrial activity such asmetformin.

A compound of Formula (I) as defined herein may also be used toadvantage in combination with known therapeutic processes, e.g., theadministration of hormones or tumor cell damaging approaches, especiallyionizing radiation.

A compound of Formula (I) as defined herein may also be used as aradiosensitizer, including, for example, the treatment of tumors whichexhibit poor sensitivity to radiotherapy.

By the term “combination”, is meant either a fixed combination in onedosage unit form, or a kit of parts for the combined administrationwhere a compound of Formula (I) and a combination partner may beadministered independently at the same time or separately within timeintervals that especially allow that the combination partners show acooperative, e.g., synergistic, effect, or any combination thereof.

The phrase, “aromatase inhibitor” as used herein relates to a compoundwhich inhibits the estrogen production, i.e., the conversion of thesubstrates androstenedione and testosterone to estrone and estradiol,respectively. The term includes, but is not limited to steroids,especially atamestane, exemestane and formestane and, in particular,non-steroids, especially aminoglutethimide, roglethimide,pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole,fadrozole, anastrozole and letrozole. Exemestane can be administered,e.g., in the form as it is marketed, e.g., under the trademark AROMASIN.Formestane can be administered, e.g., in the form as it is marketed,e.g., under the trademark LENTARON. Fadrozole can be administered, e.g.,in the form as it is marketed, e.g., under the trademark AFEMA.Anastrozole can be administered, e.g., in the form as it is marketed,e.g., under the trademark ARIMIDEX. Letrozole can be administered, e.g.,in the form as it is marketed, e.g., under the trademark FEMARA orFEMAR. Aminoglutethimide can be administered, e.g., in the form as it ismarketed, e.g., under the trademark ORIMETEN. A combination of theinvention comprising a chemotherapeutic agent which is an aromataseinhibitor is particularly useful for the treatment of hormone receptorpositive tumors, e.g., breast tumors.

The term “antiestrogen” as used herein relates to a compound thatantagonizes the effect of estrogens at the estrogen receptor level. Theterm includes, but is not limited to tamoxifen, fulvestrant, raloxifeneand raloxifene hydrochloride. Tamoxifen can be administered, e.g., inthe form as it is marketed, e.g., under the trademark NOLVADEX.Raloxifene hydrochloride can be administered, e.g., in the form as it ismarketed, e.g., under the trademark EVISTA. Fulvestrant can beformulated as disclosed in U.S. Pat. No. 4,659,516 or it can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark FASLODEX. A combination of the invention comprising achemotherapeutic agent which is an antiestrogen is particularly usefulfor the treatment of estrogen receptor positive tumors, e.g., breasttumors.

The term “anti-androgen” as used herein relates to any substance whichis capable of inhibiting the biological effects of androgenic hormonesand includes, but is not limited to, bicalutamide (CASODEX), which canbe formulated, e.g., as disclosed in U.S. Pat. No. 4,636,505.

The phrase, “gonadorelin agonist” as used herein includes, but is notlimited to abarelix, goserelin and goserelin acetate. Goserelin isdisclosed in U.S. Pat. No. 4,100,274 and can be administered, e.g., inthe form as it is marketed, e.g., under the trademark ZOLADEX. Abarelixcan be formulated, e.g., as disclosed in U.S. Pat. No. 5,843,901.

The phrase, “topoisomerase I inhibitor” as used herein includes, but isnot limited to topotecan, gimatecan, irinotecan, camptothecan and itsanalogues, 9-nitrocamptothecin and the macromolecular camptothecinconjugate PNU-166148 (compound AI in WO99/17804). Irinotecan can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark CAMPTOSAR. Topotecan can be administered, e.g., in the form asit is marketed, e.g., under the trademark HYCAMTIN.

The phrase, “topoisomerase II inhibitor” as used herein includes, but isnot limited to the anthracyclines such as doxorubicin (includingliposomal formulation, e.g., CAELYX), daunorubicin, epirubicin,idarubicin and nemorubicin, the anthraquinones mitoxantrone andlosoxantrone, and the podophyllotoxins etoposide and teniposide.Etoposide can be administered, e.g., in the form as it is marketed,e.g., under the trademark ETOPOPHOS. Teniposide can be administered,e.g., in the form as it is marketed, e.g., under the trademark VM26-BRISTOL. Doxorubicin can be administered, e.g., in the form as it ismarketed, e.g., under the trademark ADRIBLASTIN or ADRIAMYCIN.Epirubicin can be administered, e.g., in the form as it is marketed,e.g., under the trademark FARMORUBICIN. Idarubicin can be administered,e.g., in the form as it is marketed, e.g., under the trademark ZAVEDOS.Mitoxantrone can be administered, e.g., in the form as it is marketed,e.g., under the trademark NOVANTRON.

The phrase, “microtubule active agent” relates to microtubulestabilizing, microtubule destabilizing agents and microtublinpolymerization inhibitors including, but not limited to taxanes, e.g.,paclitaxel and docetaxel, vinca alkaloids, e.g., vinblastine, includingvinblastine sulfate, vincristine including vincristine sulfate, andvinorelbine, discodermolides, cochicine and epothilones and derivativesthereof, e.g., epothilone B or D or derivatives thereof. Paclitaxel maybe administered e.g., in the fo[pi]n as it is marketed, e.g., TAXOL.Docetaxel can be administered, e.g., in the form as it is marketed,e.g., under the trademark TAXOTERE. Vinblastine sulfate can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark VINBLASTIN R.P. Vincristine sulfate can be administered, e.g.,in the form as it is marketed, e.g., under the trademark FARMISTIN.Discodermolide can be obtained, e.g., as disclosed in U.S. Pat. No.5,010,099. Also included are Epothilone derivatives which are disclosedin WO 98/10121, U.S. Pat. No. 6,194,181, WO 98/25929, WO 98/08849, WO99/43653, WO 98/22461 and WO 00/31247. Included are Epothilone A and/orB.

The phrase, “alkylating agent” as used herein includes, but is notlimited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNUor Gliadel). Cyclophosphamide can be administered, e.g., in the form asit is marketed, e.g., under the trademark CYCLOSTIN. Ifosfamide can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark HOLOXAN.

The phrase, “histone deacetylase inhibitors” or “HDAC inhibitors”relates to compounds which inhibit at least one example of the class ofenzymes known as a histone deacetylase, and which compounds generallypossess antiproliferative activity. Previously disclosed HDAC inhibitorsinclude compounds disclosed in, e.g., WO 02/22577, includingN-hydroxy-3-[4-{[(2-hydroxyethyl)[2-(IH-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide,N-hydroxy-3-[4-[[[2-(2-methyl-IH-indol-3-yl)-ethylJ-amino]methyl]phenyl]-2E-2-propenamideand pharmaceutically acceptable salts thereof. It further includesSuberoylanilide hydroxamic acid (SAHA). Other publicly disclosed HDACinhibitors include butyric acid and its derivatives, including sodiumphenylbutyrate, thalidomide, trichostatin A and trapoxin.

The term “antineoplastic antimetabolite” includes, but is not limitedto, 5-Fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylatingagents, such as 5-azacytidine and decitabine, methotrexate andedatrexate, and folic acid antagonists such as pemetrexed. Capecitabinecan be administered, e.g., in the form as it is marketed, e.g., underthe trademark XELODA. Gemcitabine can be administered, e.g., in the formas it is marketed, e.g., under the trademark GEMZAR. Also included isthe monoclonal antibody trastuzumab which can be administered, e.g., inthe form as it is marketed, e.g., under the trademark HERCEPTIN.

The phrase, “platin compound” as used herein includes, but is notlimited to, carboplatin, cis-platin, cisplatinum and oxaliplatin.Carboplatin can be administered, e.g., in the form as it is marketed,e.g., under the trademark CARBOPLAT. Oxaliplatin can be administered,e.g., in the form as it is marketed, e.g., under the trademark ELOXATIN.

The phrase, “compounds targeting/decreasing a protein tyrosine or serineor threonine kinase activity” as used herein includes, but is notlimited to, gefinitib, erlotinib, lapatinib, foretinib, cabozantinib,vemurafenib or selumetinib (AZD6244). Gefinitib can be administered,e.g., in the form as it is marketed, e.g., under the trademark IRESSA.Erlotinib can be administered, e.g., in the form as it is marketed,e.g., under the trademark TARCEVA. Lapatinib can be administered, e.g.,in the form as it is marketed, e.g., under the trademarks TYKERB andTYVERB. Cabozantinib can be administered, e.g., in the form as it ismarketed, e.g., under the trademark COMETRIQ. Vemurafenib can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark CELBORAF. Foretinib can be formulated, e.g., as disclosed inUS 20,120,282,179. Selumetinib (AZD6244) can be formulated, e.g., asdisclosed in US 20,080,177,082 and US 20,090,246,274. Other suitableprotein kinase inhibitors include without limitation Afatanib (Gilotrif,Boeringer Ingelheim), Axitinib (Inlyta, Pfizer), Bosutinib (Bosulif,Wyeth), Crizotinib (Xalkori, Pfizer), Dabrafenib (Tafinlar, GSK),Dasatinib (Sprycel, Bristol-Myers Squib), Elotinib (Tarceva, OSI),Everolimus (Afinitor, Novartis), Gefitinib (Iressa, Astrazeneca),Ibrutinib (Imbruvica, Pharmacyclics and J&J), Imatanib (Gleevec,Novartis), Nilotinib (Tasigna, Novartis), Pazopanib (Votrient,GlaxoSmithKline), Ponatinib (Iclusig, Ariad), Regorafenib (Stivarga,Bayer), Ruxolitinib (Jakafi, Incyte), Sirolimus (Rapamune, Wyeth),Sorafenib (Nexavar, Bayer), Sunitinib (Sutent, Pfizer), Tofacitinib(Xeljanz, Pfizer), Temsirolimus (Torisel, Wyeth), Trametinib (Mekinist,GSK), Vandetanib (Caprelsa, IPR Pharms) as well as other proposedprotein kinase inhibitors that can be found in the literature.

Tumor cell damaging approaches refer to approaches such as ionizingradiation. The phrase, “ionizing radiation” referred to above andhereinafter means ionizing radiation that occurs as eitherelectromagnetic rays (such as X-rays and gamma rays) or particles (suchas alpha and beta particles). Ionizing radiation is provided in, but notlimited to, radiation therapy and is known in the art. See, e.g.,Hellman, Principles of Radiation Therapy, Cancer, in Principles andPractice of Oncology, Devita et al., Eds., 4th Edition, Vol. 1, pp.248-275 (1993).

The phrase, “angiostatic steroids” as used herein refers to agents whichblock or inhibit angiogenesis, such as, e.g., anecortave, triamcinolone,hydrocortisone, 11-[alpha]-epihydrocotisol, cortexolone,17[alpha]-hydroxyprogesterone, corticosterone, desoxycorticosterone,testosterone, estrone and dexamethasone.

Other chemotherapeutic agents include, but are not limited to, plantalkaloids, hormonal agents and antagonists; biological responsemodifiers, preferably lymphokines or interferons; antisenseoligonucleotides or oligonucleotide derivatives; or miscellaneous agentsor agents with other or unknown mechanism of action.

The structure of the active agents identified by code numbers, genericor trade names may be taken from the actual edition of the standardcompendium “The Merck Index” or from databases, e.g., PatentsInternational (e.g., IMS World Publications).

The above-mentioned compounds, which can be used in combination with acompound of Formula (I), can be prepared and administered as describedin the art such as in the documents cited above.

Furthermore, the compounds of the invention may be used in a method ofprofiling the functional and structural similarity of histonedemethylases comprising taking a panel of at least two histonedemethylases and a panel of at least two compounds of formula 1 anddetermining the extent to which each said compound of formula 1 inhibitsthe activity of each of said histone demethylases, and generating asimilarity index reflecting the degree of similarity between the histonedemethylases in respect of their inhibition by said compounds.

Preparation of Compounds of Formula (I): Q is CH₂NHR¹³

Method A—Reductive Amination

Compounds of Formula (I) may be prepared from 4-formyl pyridinesaccording to Scheme 1, where R′ is a suitable protecting group or R¹, inone-pot or by a stepwise procedure by mixing with an amine, optionallycontaining orthogonal protected reactive sites, and a reducing agentsuch as NaBH₄, NaBH(OAc)₃, NaCNBH₃, or Et₃SiH, either at roomtemperature or by heating for up to several hours by use of a solventsuch as an alcohol, DCE, DCM, water, or toluene, optionally adding acatalyst such as an acid or a Lewis acid. Optionally, protecting groupsmay be removed and a purification method such as silica gelchromatography is employed if needed.

Method B

Compounds of Formula (I) may be prepared from hydroxyl amines,optionally containing orthogonally protected reactive sites, accordingto Scheme 2, where R′ is a suitable protecting group or R¹, by use ofreducing agents, such as a hydrogen atmosphere over a suitable catalyst,such as palladium on charcoal, in a suitable solvent, such as analcohol. Optionally, protecting groups may be removed and a purificationmethod such as silica gel chromatography is employed if needed.

Method C

Compounds of Formula (I) may be prepared from 2-formyl pyridinesaccording to Scheme 3 analogously to Method A.

Method D—Buchwald Coupling to Aryls

Compounds of Formula (I) may be prepared according to Scheme 4 using asuitable solvent such as toluene or tetrahydrofuran, a base such ascesium carbonate or potassium t-butoxide, a suitable catalyst such asPd₂(dba)₃, optionally a suitable salt such as lithium chloride and thedesired electrophile such as arylbromide or heteroarylbromide. Thecompounds of Formula I are generated at room temperature or by heatingfor several hours, such as for 2 to 5 hours. Optionally, protectinggroups may be removed and a purification method such as silica gelchromatography is employed if needed.

Method E—Reductive Amination

Compounds of Formula (I) may be prepared from amines according to Scheme4 according to method A.

Method F—Alkylation/Acylation

The compounds of Formula (I) may be prepared according to scheme 4 byuse of a solvent such as DMF or THF, a base such as sodium hydride orcesium carbonate and a suitable electrophilic species such as anepoxide, a heteroaromatic chloride, an aliphatic, allylic or benzylicbromide, chloride or sulfonate, or a carbonyl chloride. Optionally,protecting groups may be removed and a purification method such assilica gel chromatography is employed if needed.

Method G

Compounds of Formula (I) may be prepared from amides, optionallycontaining orthogonal protected reactive sites, according to Scheme 5,where R′ is a suitable protecting group or R₁, by use of reducingagents, such as lithium aluminium hydride or borane-complexes, in asuitable solvent, such as an ether or tetrahydrofuran. Optionally,protecting groups may be removed and a purification method such assilica gel chromatography is employed if needed.

Preparation of Compounds of Formula (I): Q is CH═NR¹²

Method H

Compounds of Formula (I) may be prepared from 4-formyl pyridinesaccording to Scheme 6, where R′ is a suitable protecting group or R1, bymixing with an amine, optionally containing orthogonally protectedreactive sites, either at room temperature or by heating for up toseveral hours by use of a solvent such as an alcohol, DCE, DCM, water,or toluene, optionally adding a catalyst such as a Lewis acid.Optionally, protecting groups may be removed and a purification methodsuch as silica gel chromatography is employed if needed.

Preparation of Compounds of Formula (I): O is CH═O

Method I

Compounds of General Formula (I) may be prepared according to Scheme 7,where R′ is a suitable protecting group or R1, by a Swern oralternatively a Dess-Martin oxidation of alcohol to aldehyde.Optionally, protecting groups may be removed and a purification methodsuch as silica gel chromatography is employed if needed.

Method J

Compounds of General Formula (I) may be prepared from esters, where R′is a suitable protecting group or R1, optionally containing orthogonalprotected reactive sites, according to Scheme 8, by use of reducingagents, such as DIBAL-H, in a suitable solvent, such as toluene.Optionally, protecting groups may be removed and a purification methodsuch as silica gel chromatography is employed if needed.

Method K

Compounds of General Formula (I) may prepared at low temperature, e.g.at −78° C., from halides, where R′ is a suitable protecting group or R1,optionally containing orthogonal protected reactive sites according toScheme 9 (X designates a halogen atom) by halogen metal exchange, e.g.by treatment with an alkyl lithium reagent, followed by addition of DMFin a solvent, such as dichloromethane. Optionally, protecting groups maybe removed and a purification method such as silica gel chromatographyis employed if needed.

Preparation of Compounds of Formula (I): O is CH(OR¹⁷)₂

Method L

Compounds of General Formula (I) may prepared from 4-formyl pyridinesaccording to Scheme 10 by stirring in an alcohol in the presence of aLewis acid or an acid, such as HCL or Pyridinium toluene-4-sulphonate,optionally by reacting with trialkyl orthoformate or in the presence ofa drying agent such as an inorganic dry salt, or with azeotropic removalof water, at room temperature or by heating for several hours dependingon the method. Optionally, protecting groups may be removed and apurification method such as silica gel chromatography is employed ifneeded.

Preparation of Compounds of Formula (I): Q is W and R¹⁶ is H

Method M

Compounds of General Formula (I) may prepared from 4-formyl pyridinesaccording to Scheme 10 by stirring in a diamine, an aminoalcohol or anaminothiol, optionally in the presence of an acid such as HCL orPyridinium toluene-4-sulphonate, optionally in the presence of a dryingagent such as an inorganic dry salt or molecular sieves, or withazeotropic removal of water, at room temperature or by heating forseveral hours depending on the method. Optionally, protecting groups maybe removed and a purification method such as silica gel chromatographyis employed if needed.

Preparation of Compounds of Formula (I): Q is W and R³ is not H

Method N

Compounds of General Formula (I) may prepared from the aforementionedcompound where Q is W and R¹⁶ is H, by reacting with a suitablyactivated acyl group such as an acyl halide or acyl anhydride at roomtemperature or by heating for several hours is a solvent such asdichloroethane or THF. Optionally, protecting groups may be removed anda purification method such as silica gel chromatography is employed ifneeded.

Preparation of Intermediates for Compounds of Formula (I)

Method AA

Intermediates may be prepared from 2-formyl pyridines according toScheme 11 analogously to Method A.

Method AB

Intermediates, where X designates halides or OTf, may be prepared from2-formyl pyridines according to Scheme 12 analogously to Method A.

Method AC

Intermediates, where Pg designates a suitable protecting group, such asTBMDS or TIPS, may be prepared from 2-formyl pyridines according toScheme 13 analogously to Method A.

Method AD

Intermediates be prepared according to scheme 14, where R′ is a suitableprotecting group or R₁, by use of a solvent such as DMF or THF, a basesuch as a hindered tertiary amine, a dehydrating agent such as EDCI orDCC and an amine, and by mixing at or above room temperature for aperiod up to several hours. Optionally, the said protecting group may beremoved, and a purification method such as silica gel chromatography isemployed if needed.

Method AE

Intermediates may be prepared according to Scheme 15 analogously toMethod AD.

Method AF

Intermediates may be prepared according to Scheme 16 analogously toMethod AD.

Method AG

Intermediates may be prepared according to scheme 17 from, where R′ is asuitable protecting group or R¹ and R″ is an orthogonal protectinggroup, which may be selectively removed, such as removal of R″: ^(t)Buin presence of R′: CF₃CO by treating with trifluoroacetic acid in asolvent such as dichloromethane at room temperature for several hours. Apurification method such as silica gel chromatography is employed ifneeded.

Method AH

Intermediates may be prepared according to Scheme 18 analogously toMethod AG.

Method AI

Intermediates may be prepared according to Scheme 19 analogously toMethod AG.

Method AJ

Intermediates may be prepared from aldehydes and intermediates, where Ldesignates a bond or an aliphatic linker, which may comprise an amidebond, attached to an aliphatic heterocycle, according to Scheme 20analogously to Method A Method AK

Intermediates may be prepared according to Scheme 20 analogously toMethod F.

Method AL

Intermediates may be prepared according to Scheme 21 analogously toMethod AJ.

Method AM

Intermediates may be prepared according to Scheme 21 analogously toMethod F.

Method AN

Intermediates may be prepared according to Scheme 22 analogously toMethod AJ.

Method AO

Intermediates may be prepared according to Scheme 22 analogously toMethod F.

Method AP

Intermediates, where L designates an aliphatic linker, which maycomprise an amide bond, may be prepared from aldehydes according toScheme 23 analogously to Method E.

Method AQ

Intermediates may be prepared according to Scheme 23 analogously toMethod F.

Method AR

Intermediates, where L designates an aliphatic linker, which maycomprise an amide bond, may be prepared from aldehydes according toScheme 24 analogously to Method E.

Method AS

Intermediates may be prepared according to Scheme 24 analogously toMethod F.

Method AT

Intermediates, where L designates an aliphatic linker, which maycomprise an amide bond, may be prepared from aldehydes according toScheme 25 analogously to Method E.

Method AU

Intermediates may be prepared according to Scheme 25 analogously toMethod F.

Method AV

Intermediates may be prepared according to Scheme 26 analogously toMethod A.

Method AW

Intermediates may be prepared according to Scheme 27 analogously toMethod A.

Method AX

Intermediates may be prepared according to Scheme 28 analogously toMethod A.

Method AZ

Intermediates may be prepared from esters, optionally containingorthogonal protected reactive sites, according to Scheme 29, by use ofreducing agents, such as DIBAL-H, in a suitable solvent, such astoluene. Optionally, protecting groups may be removed and a purificationmethod such as silica gel chromatography is employed if needed.

Method BA

Intermediates may be prepared from esters, optionally containingorthogonal protected reactive sites, according to Scheme 30, by use ofreducing agents, such as lithium aluminiumhydride or borane-complexes,in a suitable solvent, such as an ether or tetrahydrofuran. Optionally,protecting groups may be removed and a purification method such assilica gel chromatography is employed if needed.

Method BB

Intermediates may be prepared according to Scheme 31 using method K

Method BC

Intermediates may be prepared according to scheme 31 either at roomtemperature or by heating for several hours by use of a solvent such astoluene or tetrahydrofuran, a base such as cesium carbonate or potassiumt-butoxide, a catalyst such as Pd₂(dba)₃, optionally a salt such aslithium chloride and the desired nucleophile such as carbon monoxide. Apurification method such as silica gel chromatography is employed ifneeded.

Method BD

Intermediates may be prepared according to scheme 32 analogously toMethod AD.

Method BE

Intermediates may be prepared according to Scheme 33 by use of a solventsuch as DMF or THF, a base such as cesium carbonate and an electrophilesuch as an alkyl halide, heteroaromatic halide, alkenyl halide, etc.,and by mixing at or above room temperature for several hours. Apurification method such as silica gel chromatography or trituration isemployed if needed.

Method BF

Intermediates may be prepared according to Scheme 33 by use of aceticcatalysis in an alcohol at room temperature or at reflux. A purificationmethod such as silica gel chromatography or trituration is employed ifneeded.

Method BG

Intermediates may be prepared according to scheme 34 from 4-formylpyridines by reaction with hydroxylamine in a solvent such as an alcoholor water.

Method BH

Intermediates may be prepared according to scheme 35 from 4-formylpyridines by with reaction an amine, optionally containing orthogonallyprotected reactive sites, either at room temperature or by heating forup to several hours by use of a solvent such as an alcohol, DCE, DCM,THF water, or toluene, optionally adding a catalyst such as a Lewisacid. Subsequently reacting with TMSCN in a solvent such asacetonitrile. Optionally, protecting groups may be removed and apurification method such as silica gel chromatography is employed ifneeded.

Method BI

Intermediates may be prepared according to Scheme 36 either at roomtemperature or by heating for several hours by use of a solvent such aswet toluene or tetrahydrofuran, a base such as cesium carbonate orpotassium t-butoxide, a catalyst such as Pd₂(dba)₃, optionally a saltsuch as lithium chloride and the desired nucleophile such as carbonmonoxide. A purification method such as silica gel chromatography isemployed if needed.

Method BJ

Intermediates may be prepared according to Scheme 37 from pyridine2-carboxylates analogously to Method J.

Method BK

Intermediates may be prepared according to Scheme 38 from pyridine2-halides analogously to Method K.

EXAMPLES Example 1 Preparation of Compounds of the Invention

General Methods and Materials

All chemicals were purchased from Sigma-Aldrich, Alfa Aesar, Matrix,Combiblock, Oakwood, and Chembridge. Anhydrous solvents were AldrichSure/Seal™ brand. All reactions were carried out under a dry nitrogenatmosphere using dry solvents. Reactions were monitored by thin-layerchromatography carried out on Sigma-Aldrich 0.25 mm silica gel plates(60 Å, fluorescent indicator). Spots were visualized under UV light (254nm). Flash column chromatography was performed on Biotage SNAP FlashSystem, or silica gel 60 (particle size 0.032-0.063 mm) obtained fromSilicycle, Inc. Low-resolution ES (electrospray) mass spectra wereobtained using a Micromass Quattro Ultima mass spectrometer in theelectrospray positive (ES+) or negative (ES−) ion mode. 1H-NMR spectrawere recorded on a Bruker AM-300 spectrometer and were calibrated usingresidual nondeuterated solvent as internal reference. Spectra wereprocessed using Spinworks version 2.5 (developed by Dr. Kirk Marat,Department of Chemistry, University of Manitoba). Preparative HPLC wasperformed on Waters 2996 with Photodiode Array Detector, Waters 600Controller, Waters 100 pump, and Waters 717 auto sampler, with UVdetection at 254 and 280 nm. Flow rate: 15 mL/minute, run time 30minutes. Solvents: 0-100% (H₂O-MeOH), with and without added TFA (0.1%).Column used was Supelco C18, 25 cm×21.2 mm, particle size 10 micrometer.

Ethyl 2-formylpyridine-4-carboxylate was prepared analogously toQueguiner, G. and Pastour, P. (Comptes Rendus des Seances de l'Académiedes Sciences, Série C: Sciences Chimiques (1969), 268(2), 182-5).

Examples of Compounds of Formula (I)

TABLE 1 May be prepared analo- gously to Synthetic Structure # NameRoute NMR

1 N-{[2-({[4- (diethylamino)butyl]- amino}methyl)pyridin-4-yl]methyl}-2,2,2- trifluoroacetamide A ¹H NMR (300 MHz, Methanol-d₄), δppm: 8.57 (d, 1H), 4.53 (s, 2H), 1.94 (s, 6H), 1.25 (t, 6H).

2 [2-({[4- (dimethylamino)butyl]- amino}methyl)pyridin- 4-yl]methanamineB ¹H-NMR (300 MHz, MeOD), δ ppm: 8.80 (d, 1H), 4.60 (s, 2H), 2.95 (s,6H), 1.95 (m, 4H)

3 [2-({[3- (dimethylamino)propyl]- amino}methyl)pyridin-4-yl]methanamine B ¹H NMR (300 MHz, Methanol-d₄), δ ppm: 8.71 (d, 1H),4.48 (s, 2H), 2.95 (s, 6H), 2.25 (m, 2H).

4 2-({[4- (aminomethyl)pyridin-2- yl]methyl}amino)-N-[2-(dimethylamino)ethyl]- N-ethylacetamide B ¹H NMR (300 MHz, CDCl₃), δppm: 8.74 (d, 1H), 4.59 (s, 2H), 2.99 (s, 6H), 1.27 (t, 3H).

5 [2-({[4- (diethylamino)butyl]- amino}methyl)pyridin-4- yl]methanamineB ¹H-NMR (300 MHz, MeOD), δ ppm: 8.82 (d, 1H), 4.65 (s, 2H), 3.20 (m,9H), 1.30 (t, 6H).

6 N-[4- (diethylamino)butyl]-- 2,2,2-trifluoro-N-({4-[(trifluoroacetamido)- methyl]pyridin-2- yl}methyl)acetamide C ¹H-NMR(300 MHz, CDCl3), δ ppm: 11.60, 11.45 (d, 1H), 4.70 (d, 2H), 3.10 (m,4H), 1.50 (t, 6H).

7 [2-({[4-(azetidin-1- yl)butyl]amino}methyl)- pyridin-4-yl]-methanamine D ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.70 (d, 1H), 4.40 (s,2H), 1.80 (m, 4H), 1.20 (m 4H).

8 [2-({[5- (dimethylamino)pentyl]- amino}methyl)pyridin-4-yl]methanamine B ¹H-NMR (300 MHz, MeOD), δ ppm: 8.88 (d, 1H), 4.70 (s,2H), 2.85 (s, 6H), 1.85 (m, 4H).

9 2-({[4- (aminomethyl)pyridin- 2-yl]methyl}amino)- N-{1-[(2-methoxyphenyl)methyl]- piperidin-4-yl}acetamide B ¹H-NMR (300 MHz,Methanol-d₄), δ ppm: 8.74 (m, 1H), 7.18- 7.10 (m, 2H), 4.45 (d, 2H),2.25-1.80 (m; 4H).

10 N-{[2-({[4- (dimethylamino)butyl]- amino}methyl)pyridin-4-yl]methyl}- cyclopropanamine E ¹H NMR (300 MHz, CDCl₃), δ ppm: 8.42 (d,1H), 2.20 (s, 6H), 2.15 (m, 1H), 0.5-0.38 (m, 4H).

11 N-{[2-({[3-(2- methylpiperidin-1- yl)propyl]amino}methyl)- pyridin-4-yl]methyl}- cyclopropanamine E ¹H-NMR (300 MHz, Methanol-d₄), δ ppm:8.80 (d, 1H), 2.90 (m, 1H), 2.30 (m, 2H), 1.02-0.9 (m, 4H).

12 N-({2- [(propylamino)methyl]- pyridin-4- yl}methyl)- cyclopropanamineE ¹H NMR (300 MHz, Methanol-d₄), δ ppm: 8.70 (d, 1H), 4.40 (s, 2H), 3.06(m, 1H), 1.03 (t, 3H).

13 2-{[(4- {[(cyanomethyl)amino]- methyl}pyridin-2-yl)methyl]amino}-N,N- dimethylacetamide F ¹H NMR (300 MHz, Methanol-d₄),δ ppm: 8.72 (d, 1H), 4.37 (s, 2H), 4.18 (s, 2H), 3.00 (s, 6H).

14 2-{[(4-{[(2- fluoroethyl)amino]- methyl}pyridin-2-yl)methyl]amino}-N,N- dimethylacetamide G ¹H NMR (300 MHz, Methanol-d₄),δ ppm: 8.49 (d, 1H), 4.65 (t, 1H), 3.97 (s, 2H), 2.96 (d, 6H).

15 2-({[4-({[2- (dimethylamino)ethyl]- amino}methyl)pyridin-2-yl]methyl}amino)-N,N- dimethylacetamide G ¹H NMR (300 MHz, Methanol-d₄),δ ppm: 8.44 (d, 1H), 3.90 (s, 2H), 2.96 (d, 6H), 2.25 (s, 6H).

16 {[(2S)-1- benzylpyrrolidin-2- yl]methyl}[(4- {[(cyclopropylmethyl)-amino]methyl}pyridin- 2-yl)methyl]amine G ¹H NMR (300 MHz, Methanol-d₄),δ ppm: 8.70 (dd, 1H), 7.59- 7.44 (m, 7H), 2.25- 2.04 (m, 3H), 1.20- 1.08(m, 1H).

17 benzyl(methyl){3-[({4- [(methylamino)methyl]- pyridin-2-yl)methyl)-amino]propyl}- amine H ¹H NMR (300 MHz, CDCl₃), δ ppm: 8.8 (d, 1H),7.8-7.4 (m, 7H), 2.8 (d, 6H), 2.5-2.2 (m, 2H).

18 benzyl[3-({[4-({[2- (dimethylamino)ethyl]- amino}methyl)pyridin-2-yl]methyl}- amino)propyl]- methylamine H ¹H NMR (300 MHz,Methanol-d₄), δ ppm: 8.8 (d, 1H), 7.6-7.4 (m, 5H), 3.05 (s, 6H), 2.8 (s,3H).

19 benzyl(3-{[(4-{[(2- methoxyethyl)amino]- methyl}pyridin-2-yl)methyl]amino}- propyl)methylamine H ¹H NMR(300 MHz, Methanol-d₄), δppm: 8.9 (d, 1H), 7.6-7.4 (m, 5H), 3.8-3.2 (m, 11H), 2.9 (s, 3H).

20 2-[({4- [(cyclopropylamino)- methyl]pyridin-2- yl}methyl)amino]-N-{1-[(2- methoxyphenyl)methyl]- piperidin-4-yl}acetamide I ¹H-NMR (300 MHz,Methanol-d₄), δ ppm: 8.78 (m, 1H), 7.02 (m, 1H), 2.60 (m, 1H), 0.9- 0.8(m, 4H).

21 2-cyclopropyl-2-({[2- ({[2- (dimethylamino)ethyl]-amino}methyl)pyridin- 4-yl]methyl}amino)- acetonitrile J ¹H NMR (300MHz, Methanol-d₄), δ ppm: 8.68 (d, 1H), 4.50 (s, 2H), 3.00 (s, 6H), 0.61(m, 2H).

22 2-({[2-({[3- (dimethylamino)propyl]- amino}methyl)pyridin-4-yl]methyl}amino)- propanenitrile J ¹H NMR(300 MHz, Methanol-d₄), δ ppm:8.61 (d, 1H), 4.43 (s, 2H), 2.94 (s, 6H), 1.54 (d, 3H).

23 2-[({2-[({4- [benzyl(cyclopropyl)- amino]butyl}amino)-methyl]pyridin-4- yl}methyl)amino]- acetonitrile K ¹H-NMR (300 MHz,CDCl₃), δ ppm: 8.45 (d, 1H), 7.20-7.00 (m, 7H), 1.40 (m, 12H) 0.40 (m,4H), ppm.

24 2-[2-({[3- (dimethylamino)propyl]- amino}methyl)pyridin-4-yl]-2-(methylamino)- acetonitrile J ¹H NMR (300 MHz, Methanol-d₄), δppm: 8.83 (d, 1H), 4.54 (s, 3H), 2.93 (s, 6H), 2.25 (m, 2H).

25 N-[(2-{[({[2- (dimethylamino)ethyl]- (ethyl)carbamoyl}-methyl)amino]methyl}- pyrtdin-4- yl)methyl]-2,2,2- trifluoroacetamide L¹H NMR (300 MHz, Methanol-d₄), δ 8.61 (d, 1H), 7.35 (d, 1H), 4.53 (s,2H), 4.46 (s, 2H), 4.23 (s, 2H), 3.82 (t, 2H), 2.98 (s, 6H), 1.24 (t,3H).

26 N-[(2-{[N-({[2- (dimethylamino)ethyl]- (ethyl)carbamoyl}-methyl)-2,2,2- trifluoroacetamido]- methyl}pyridin-4- yl)methyl]-2,2,2-trifIuoroacetamide C ¹H-NMR (300 MHz, CDCl₃), δ ppm 8.50 (m, 1H), 7.88(m, 1H), 7.21 (m, 2H), 4.53 (m, 6H), 3.33 (m 4H), 2.44 (m, 2H), 2.23 (m,6H), 1.14 (m, 3H).

27 ({[2-({[4- (diethylamino)- butyl]amino} methyl)pyridin-4-yl]methyl}carbamoyl)- formic acid M ¹H NMR(300 MHz, Methanol-d₄), δ ppm:8.40 (d, 1H), 4.45 (s, 2H), 2.70-2.40 (m, 8H), 1.00 (m, 6H).

28 tert-butyl ({[2-({[4- (diethylamino)- butyl]amino} methyl)pyridin-4-yl]methyl}carbamoyl)- formate M ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.50 (m,1H), 4.50 (m, 2H), 1.65-1.50 (m, 13H), 1.20 (m, 6H).

29 ethyl 2-({[(2-{[({[2- (dimethylamino)- ethyl](ethyl)-carbamoyl}methyl)- amino]methyl}pyridin- 4-yl)methyl]- carbamoyl}-oxy)benzoate L ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.50 (m, 1H), 7.20 (m,4H), 4.50 (m, 2H), 4.40 (q, 2H), 250 (s, 6H), 1.38 (t, 3H).

30 N-[(2-{[({[2-(azetidin- 1-yl)ethyl](ethyl)- carbamoyl}-methyl)amino]methyl}- pyridin-4-yl)methyl]- 2,2,2-trifluoroacetamide A¹H NMR (300 MHz, Methanol-d₄), δ 8.50 (d, 1H), 7.40 (m, 2H), 4.50 (s,2H), 4.40 (s, 2H), 4.20 (m, 2H), 3.50 (m, 2H), 3.30 (m, 2H) 2.40 (m,2H), 1.10 (t, 3H)

31 N-[(2-{[({[2- (dimethylamino)ethyl] (ethyl)carbamoyl}-methyl)amino]methyl}- pyridin-4-yl)- methyl]-2,2,3,3,4,4,4-heptafluorobutanamide L ¹H NMR (300 MHz, Methanol-d₄), δ ppm: 8.70, (d,1H), 4.51(s, 2H), 2.98 (s, 6H), 1.22 (m, 3H).

32 N-[(2-{[({[2- (dimethylamino)- ethyl](ethyl)- carbamoyl}methyl)-amino]methyl}pyridin- 4-yl)methyl]-2,2- difluorobutanamide L ¹H NMR (300MHz, Methanol-d₄), δ 8.60 (d, 1H), 7.39 (s, 1H), 7.36 (d, 1H), 4.48 (m,4H), 4.24 (s, 2H), 3.82 (t, 2H), 3.38 (m, 4H), 2.98 (s, 6H), 2.32 (m,2H), 1.24 (t, 3H), 1.02 (t, 3H).

33 2-[({4-[(N- cyclopropyl- carboximidoyl] pyridin-2-yl}methyl)amino]-N,N- dimethylacetamide N ¹H NMR (300 MHz, CDCl₃), δppm: 8.55 (d, 1H), 8.52 (s, 1H), 3.17 (m, 1H), 2.96 (d, 6H).

34 N,N-dimethyl-2-[({4- [[(3- phenylpropyl)imino]- methyl]pyridin-2-yl}methyl)amino]- acetamide N ¹H NMR (300 MHz, CDCl₃), δ ppm: 8.62 (d,1H), 8.26 (s, 1H), 7.25 (m, 5H), 2.96 (d, 6H).

35 N,N-dimethyl-2- [({4-[N-(2- methylcyclopropyl)-carboximidoyl]pyridin- 2-yl}methyl)amino]- acetamide O ¹H NMR (300 MHz,CDCl₃), δ ppm: 8.57 (d, 1H), 8.36 (s, 1H), 2.96 (d, 6H), 0.81 (m, 1H).

36 2-[({4-[[(2- cyclohexylethyl)imino]- methyl]pyridin-2-yl}methyl)amino]-N,N- dimethylacetamide O ¹H NMR (300 MHz, CDCl₃), δppm: 8.61 (d, 1H), 8.26 (s, 1H), 2.95 (d, 6H), 1.78-0.88 (m, 13H).

37 [3- (dimethylamino)propyl]- ({4-[{[3- (dimethylamino)propyl]-imino}methyl]pyridin-2- yl}methyl)amine N ¹H NMR (300 MHz, CDCl₃), δppm: 8.59 (dd, 1H), 8.24 (s, 1H), 2.20 (s, 6H), 2.19 (s, 6H).

38 ({4-[{[2- (dimethylamino)ethyl]- imino}methyl]pyridin-2-yl}methyl)[3- (dimethylamino)- propyl]amine N ¹H NMR (300 MHz, CDCl₃), δppm: 8.60 (d, 1H), 8.28 (s, 1H), 2.30 (s, 6H), 2.25 (s, 6H).

39 N-{[2-({[2- (ethylsulfanyl)- ethyl]amino} methyl)pyridin-4-yl]methylidene}- cyclopropanamine P ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.5(d, 1H), 8.4 (s, 1H), 1.2 (t, 3H), 1.0 (m, 4H).

40 N-{[2-({[2-(l- methylpyrrolidin-2- yl)ethyl]amino}methyl)- pyridin-4-yl]methylidene}- cyclopropanamine P ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.5(d, 1H), 8.4 (s, 1H), 2.8 (s, 3H), 1.0 (m, 4H).

41 N-({2-[({3- [benzyl(methyl)amino]- propyl}amino)methyl]- pyridin-4-yl}methylidene)- cyclopropanamine Q ¹H NMR (300 MHz, CDCl3), 

 ppm: 8.55 (d, 1H), 8.50 (s, 1H), 7.28-7.22 (m, 5H), 1.04-0.97 (m, 4H).

42 N-{[2-({[3-(pyrrolidin- 1-yl)propyl]amino}- methyl)pyridin-4-yl]methylidene}- cyclopropanamine P ¹H-NMR (300 MHz, Methanol-d₄), δppm: 8.58 (m, 1H), 8.40 (s, 1H), 2.78-2.60 (m, 6H), 1.90-1.75 (m, 6H).

43 N-{[2-({[(2E)-4- (dimethylamino)but-2- en-1-yl]amino}-methyl)pyridin- 4-yl]methylidene}- cyclopropanamine R ¹H NMR (300 MHz,CDCl₃), δ ppm: 8.59 (d, 1H), 8.41 (s, 1H), 2.23 (s, 6H), 0.94 (m, 4H).

44 N-{[2-({[4-(azetidin-1- yl)butyl]amino}methyl)- pyridin-4-yl]methylidene}- cyclopropanamine S ¹H-NMR (300 MHz, CDCl₃), ), δ ppm:8.50 (d, 1H), 8.40 (s, 1H), 1.50 (m 6H), 1.00 (m, 4H).

45 N-{[2-({[4- (dimethylamino)butyl]- amino}methyl)- pyridin-4-yl]methylidene}- cyclopropanamine T ¹H-NMR (300 MHz, CDCl3), ), δ ppm:8.50 (s, 1H), 8.30 (s, 1H), 2.20 (s, 6H), 1.00 (m, 4H)

46 N-[(2-{[({4- [(dimethylamino)- methyl]cyclohexyl}- methyl)amino]-methyl}pyridin-4- yl)methylidene]- cyclopropanamine N ¹H NMR (300 MHz,CDCl₃), δ ppm: 8.57 (d, 1H), 8.41 (s, 1H), 2.20 (s, 6H), 0.98 (m, 8H).

47 N-{[2-({[5- (dimethylamino)- pentyl]amino}- methyl)pyridin-4-yl]methylidene}- cyclopropanamine U ¹H-NMR (300 MHz, CDCl3), ), δ ppm:8.50 (d, 1H), 8.40 (s, 1H), 2.15 (s, 6H), 1.50-1.30 (m, 6H).

48 2-[({4-[N- cyclopropyl- carboximidoyl] pyridin-2- yl}methyl)amino]-N-[4-(diethylamino)- butyl]acetamide V ¹H NMR (300 MHz, CDCl₃), δ ppm:8.61 (d, 1H), 8.41 (s, 1H), 1.09 (t, 6H), 1.03 (m, 4H).

49 2-[({4-[N- cyclopropyl- carboximidoyl] pyridin-2- yl}methyl)amino]-1-[(2R)-2-(pyrrolidin-1- ylmethyl)pyrrolidin-1- yl]ethan-1-one V ¹H NMR(300 MHz, CDCl₃), δ ppm: 8.57 (d, 1H), 8.41 (s, 1H), 1.81- 1.69 (m, 4H),1.07 (m, 4H).

50 N-(2-cyanoethyl)- 2-[({4-[N- cyclopropyl- carboximidoyl]- pyridin-2-yl}methyl)amino]-N- ethylacetamide V ¹H NMR (300 MHz, CDCl₃), δ ppm:8.56 (d, 1H), 8.40 (s, 1H), 1.17 (t, 3H), 1.04-0.95 (m, 4H).

51 2-[({4-[N- cyclopropyl- carboximidoyl]- pyridin-2- yl}methyl)-amino]-N-[(1- ethylpyrrolidin-2- yl)methyl]acetamide V ¹H-NMR (300 MHz,CDCl3), ), δ ppm: 8.50 (d, 1H), 8.40 (s, 1H), 1.70 (s, 1H), 1.10 (m,6H).

52 2-[({4-[N- cyclopropyl- carboximidoyl]- pyridin-2-yl}methyl)amino]-N- methyl-N-[3-(1H- pyrazol-1- yl)propyl]acetamide V ¹HNMR (300 MHz, CDCl₃), δ ppm: 8.57 (d, 1H), 8.41 (s, 1H), 6.22 (m, 1H),1.01 (m, 4H).

53 N-(1-benzylpyrrolidin- 3-yl)-2-[({4-[N- cyclopropyl- carboximidoyl]pyridin-2- yl}methyl)amino]- acetamide V ¹H-NMR (300 MHz, CDCl3), ), δppm: 8.50 (d, 1H), 8.35 (s, 1H), 7.20 (m, 6H), 1.60 (m, 1H).

54 2-[({4-[N- cyclopropyl- carboximidoyl]- pyridin-2-yl}methyl)amino]-1- (4-methylpiperazin-1- yl)ethan-1-one V ¹H NMR (300MHz, CDCl₃), δ ppm: 8.56 (d, 1H), 8.39 (s, 1H), 2.29 (s, 3H), 0.99 (m,4H).

55 1-(4-benzylpiperidin- 1-yl)-2-[({4-[N- cyclopropyl- carboximidoyl]-pyridin-2- yl}methyl)amino]- ethan-1-one V ¹H NMR (300 MHz, CDCl₃), δppm: 8.59 (d, 1H), 8.42 (s, 1H), 7.32- 7.12 (m, 5H), 0.99 (m, 4H).

56 2-[({4-[N- cyclopropyl- carboximidoyl]- pyridin-2-yl}methyl)amino]-N- methyl-N-(prop-2-yn-1- yl)acetamide V ¹H NMR (300MHz, CDCl₃), δ ppm: 8.58 (d, 1H), 8.42 (s, 1H), 3.01 (m, 3H), 1.02 (m,4H).

57 2-[({4-[[(2- cyclohexylethyl)imino]- methyl]pyridin-2-yl}methyl)amino]-N,N- diethylacetamide V ¹H-NMR (300 MHz, CDCl₃), δ ppm:8.6 (d, 1H), 8.3 (s, 1H), 1.6 (m, 6H), 1.4-1.1 (m, 11H)

58 N,N-diethyl-2-[({4- [(octylimino)methyl]- pyridin-2-yl}methyl)amino]- acetamide V ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.6 (d,1H), 8.2 (s, 1H), 1.4 (m, 10H), 0.9 (t, 3H)

59 methyl 2-[({4-[N- cyclopropyl- carboximidoyl] pyridin-2-yl}methyl)amino]- acetate P ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.6 (d, 1H),8.3 (s, 1H), 3.7 (s, 3H), 3.1 (m, 1H)

60 [4- (diethylamino)butyl]- ({4-[[(2- methoxyethyl)imino]-methyl]pyridin-2- yl}methyl)amine X ¹H NMR (300 MHz, CDCl₃), δ ppm: 8.61(d, 1H), 8.29 (s, 1H), 2.51 (q, 4H), 1.01 (t, 6H).

61 2-[{[2-({[4- (diethylamino)- butyl]amino}- methyl)pyridin-4-yl]methylidene}- amino]ethan-1-ol N ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.6(d, 1H), 8.3 (s, 1H), 1.5 (m, 4H), 1.0 (t, 6H)

62 {[2-({[4- (diethylamino)- butyl]amino} methyl)pyridin-4-yl]methylidene} (2,2,3,3,3- pentafluoropropyl)- amine N ¹H-NMR (300 MHz,CDCl3), δ ppm: 8.53 (d, 1H), 8.30 (s, 1H), 1.90 (m, 4H), 1.30 (t, 6H).

63 2-[({4-[[(2- cydohexylethyl)- imino] methyl]pyridin-2-yl}methyl)amino]- N-[2-(dimethylamino)- ethyl]-N- ethylacetamide X ¹HNMR (300 MHz, CDCl₃), δ ppm: 8.56 (d, 1H), 8.22 (s, 1H), 1.76- 1.51 (m,7H), 1.22-1.07 (m, 6H)

64 [3- (dimethylamino)- propyl]({4- [(methoxyimino)- methyl]pyridin-2-yl}methyl)amine N ¹H NMR (300 MHz, Methanol-d₄), δ ppm: 8.66 (d, 1H),8.15 (s, 1H), 4.01 (s, 3H), 2.93 (s, 6H).

65 [4-(diethylamino)- butyl]({[4- (1-methylimidazolidin- 2-yl)pyridin-2-yl]methyl})amine N ¹H-NMR (300 MHz, CDCl₃), δ ppm: 8.5 (d, 1H), 7.4 (s,1H), 2.3 (s, 3H), 1.0 (t, 6H)

66 N-[2-(dimethylamino) ethyl]-N- ethyl-2-[({4-[([(2-hydroxyethyl)imino]- methyl]pyridin-2- yl}methyl)amino]- acetamide X 1HNMR (300 MHz, CDCl3): δ ppm 8.54 (d, 1H), 3.93 (s, 2H), 3.43 (s, 2H),3.40-3.32 (m, 2H), 3.25-3.17 (m, 2H),, 2.21 and 2.16 (two singlets, 6H,rotamers).

67 (2-cyclohexylethyl)- ({[2-({[4- (diethylamino)- butyl]amino}methyl)pyridin-4- yl]methylidene})- amine X 1H NMR (300 MHz, CDCl3) δppm 8.56 (d, 1H), ,3.93 (s, 2H), , 2.65 (q, 4H), 2.56 (t, 2H), 1.05 (t,6H)

68 [4- (diethylamino)butyl]({[4- (1-methyl-1,3-diazinan- 2-yl)pyridin-2-yl]methyl})amine N 1H NMR (300 MHz, chloroform-d): δ ppm 8.49 (d, 1H),3.87 (s, 2H), 3.68 (s, 1H), 3.51 (q, 4H), 2.36 (m, 4H), 1.93 (s, 3H),0.99 (t, 6H).

69 N,N-diethyl-2- [({4-[{[2-(4- methylphenyl)- ethyl]imino}-methyl]pyridin-2- yl}methyl)amino]- acetamide O 1H NMR (300 MHz, CDCl3),δ ppm: 8.6 (d, 1H), 8.1 (s, 1H), 4.1 (s, 2H), 3.8 (m, 2H), 3.6 (s, 2H),1.1 (m, 6H).

70 4-[2-{[2-({[4- (diethylamino)- butyl]amino} methyl)pyridin-4-yl]methylidene}- hydrazin- 1-yl]benzonitrile N 1H NMR (300 MHz,chloroform-d): δ ppm 8.63 (d, 1H), 7.88 (s, 1H), 7.82 (s, 1H), 4.46 (s,2H), 3.32 (m, 4H), 1.34 (t, 6H).

71 3-[{[2-({[4- (diethylamino)- butyl]amino} methyl)pyridin-4-yl]methylidene}amino]- propan-1-ol X 1H NMR (300 MHz, chloroform-d): δppm 8.61 (d, 1H), 7.88 (s, 1H), , 2.67 (m, 2H), 2.51 (q, 4H), 1.00 (t,6H).

72 [4-(diethylamino)- butyl][(4-{7-oxa-9- azaspiro[4.5]decan-8-yl}pyridin-2- yl)methyl]amine X 1H NMR (300 MHz, chloroform-d): δ ppm8.53 (d, 1H), 7.44 (s, 1H), 5.08 (s, 1H), 2.53 (q, 4H), 1.57 (m, 8H),1.02 (t, 6H).

73 2-[{[2-({[4- (diethylamino)- butyl]amino}- methyl)pyridin-4-yl]methylidene}- amino]propan-1-ol X 1H NMR (300 MHz, chloroform-d): δppm 8.60 (d, 1H), 8.32 (s, 1H), 3.94 (s, 2H), , 2.51 (q, 4H), 1.24 (d,3H), 1.02 (t, 6H).

74 1-[{[2-({[4- (diethylamino)- butyl]amino}- methyl)pyridin-4-yl]methylidene}- amino]propan-2-ol X 1H NMR (300 MHz, chloroform-d): δppm 8.62 (d, 1H), 8.30 (s, 1H), 4.11 (m, 1H), 3.95 (s, 2H), 2.52 (q,4H), 1.01 (t, 6H).

75 2-[{[2-({[4- (diethylamino)- butyl]amino}- methyl)pyridin-4-yl]methylidene}- amino]-2- phenylethan-1-ol X 1H NMR (300 MHz,chloroform-d): δ ppm 8.57 (d, 1H), 8.36 (s, 1H), 7.37 (m, 6H), 4.51 (m,1H), 2.51 (q, 4H), 1.00 (t, 6H).

76 3-[{[2-({[4- (diethylamino)- butyl]amino}- methyl)pyridin-4-yl]methylidene}- amino]-2,2- dimethylpropan-1-ol X 1H NMR (300 MHz,chloroform-d): δ ppm 8.54 (d, 1H), 7.45 (s, 1h), 3.91 (s, 2H), 2.52 (q,4H), 1.00 (t, 6H), 0.96 (s, 6H).

77 (1-{[{[2-({[4- (diethylamino)- butyl]amino}- methyl)pyridin-4-yl]-methylidene}amino]- methyl}cyclopropyl)- methanol X 1H NMR (300 MHz,chloroform-d): δ ppm 8.64 (d, 1H), 8.19 (s, 1H), 3.94 (s, 2H), 2.53 (q,4H), 1.02 (t, 6H), 0.52 (m, 4H).

78 N-[2-(dimethylamino)- ethyl]-N- ethyl-2-[({4-[[(3-hydroxypropyl)imino]- methyl]pyridin-2-yl}- methyl)amino]- acetamide X1H NMR (300 MHz, chloroform-d): δ: 8.6 (m, 1H), 8.2 (s, 1H), 3.5 (m,2H), 2.3 (s, 3H), 2.2 (s, 3H), 1.1 (m, 3H).

79 N-ethyl-2-[({4-[[(2- hydroxyethyl)imino]- methyl]pyridin-2-yl}methyl)amino]-N-[(1- methylpyrrolidin-2-yl)- methyl]acetamide X 1H NMR(300 MHz, CDCl3): δ ppm 8.59 (d, 1H), 8.31 (s, 1H), 3.98 (s, 2H), , 2.44(m, 1H), 2.37 and 2.28 (two singlets, 3H), , 1.16-1.10 (m, 3H).

80 2-{[{[2-({[4- (diethylamino)- butyl]amino} methyl)pyridin-4-yl]methylidene}- amino]methyl}-3- phenylpropan-1-ol X 1H NMR (300 MHz,chloroform-d): δ ppm 8.51 (d, 1H), 7.44 (s, 1H), 7.24 (m, 6H), 3.94 (s,2H), 3.70 (m, 2H), 2.72 (m, 2H), 2.61 (q, 4H), 2.49 (m, 4H), 1.57 (m,4H), 1.06 (t, 6H).

81 2-[({4-[[(2- cyclohexyl-3- hydroxypropyl)- imino]methyl]pyridin-2-yl}methyl)- amino]-N-[2- (dimethylamino)- ethyl]-N-ethylacetamide X 1H NMR (300 MHz, chloroform-d): δ ppm: 8.6 (m, 1H), 8.2(s, 1H), 4.0-3.8 (m, 4H), 2.3 (s, 3H), 2.2 (s, 3H), 1.8-1.6 (m, 12H),1.1 (m, 3H).

82 N-[3- (dimethylamino)propyl]- N-ethyl-2-[({4-[[(2-hydroxyethyl)imino]- methyl]pyridin-2- yl}methyl)amino]- acetamide X 1HNMR (300 MHz, CDCl3): δ ppm 8.55 (d, 1H), 8.27 (s, 1H), 3.95 (s, 2H),,2.18 and 2.13 (two singlets, 6H), 1.13- 1.07 (m, 3H).

83 N-[2-(dimethylamino)- propyl]- N-ethyl-2-[({4-[[(2-hydroxyethyl)imino]- methyl]pyridin-2- yl}methyl)amino]- acetamide O 1HNMR (300 MHz, CDCl3): δ ppm 8.59 (d, 1H), 8.31 (s, 1H), 3.98 (s, 2H),3.43 (s, 2H), 3.43 (m, 1H), , 2.25 and 2.19 (two singlets, 6H), 0.91 (d,3H).

84 1-[{[2-({[4- (diethylamino)- butyl]amino} methyl)pyridin-4-yl]methylidene}amino]- 3-phenylpropan-2-ol N 1H NMR (300 MHz,chloroform-d): δ ppm 8.63 (d, 1H), 8.31 (s, 1H), 7.27 (m, 5H), 4.19 (m,1H), 3.96 (s, 2H), 2.51 (q, 4H), 1.01 (t, 6H).

85 N-{[(1S,2S)-2- (dimethylamino)- cyclopentyl]methyl}-N-ethyl-2-[({4-[[(2- hydroxyethyl)imino]- methyl]pyridin-2-yl}methyl)amino] acetamide- N 1H NMR (300 MHz, CDCl3): δ ppm 8.55 (d,1H), 8.27 (s, 1H), 3.96 (s, 2H), 2.38 (m, 1H), 2.23 and 2.16 (twosinglets, 6H), 1.65-1.41 (m, 5H), 1.12-1.07 (m, 3H).

86 2-[({4-[{[3- (dimethylamino)-2- hydroxypropyl]imino}-methyl]pyridin-2- yl}methyl)amino]- N-[2-(dimethylamino)-ethyl]-N-ethylacetamide N 1H NMR(300 MHz, chloroform-d): δ ppm: 8.6 (d,1H), 8.2 (s, 1H), 4.0-3.8 (m, 4H), 3.3 (m, 6H), 2.3-2.1 (m, 12H), 1.1(m, 3H).

87 2-({[4-(5,5-dimethyl- 1,3-oxazinan-2- yl)pyridin-2- yl]methyl}amino)-N-[2-(dimethylamino)- ethyl]-N-ethylacetamide N NMR (300 MHz,chloroform-d): δ ppm 8.50 (m, 1H), 5.00 (s, 1H), 4.00 (s, 2H), 2.20 (d,6H), 1.10 (m, 6H).

88 N-[2- (dimethylamino)- ethyl]-N- ethyl-2-[({4-[({[1- (hydroxymethyl)cyclopropyl]methyl} imino)methyl]pyridin-2- yl}methyl)amino] acetamide N1H NMR (300 MHz, CD₃OD), δ ppm: 8.9 (d, 1H), 7.7 (s, 1H), 7.6 (d, 1H),5.9 (s, 1H), 4.6 (s, 2H), 4.3, 2H), 3.8 (m, 2H), 3.5 (s, 2H), 3.3 (m,8H), 1.2 (m, 3H), 0.9 (m, 3H), 0.6 (m, 2H), 0.4 (m, 2H).

89 2-[({4-[[(2-benzyl-3- hydroxypropyl)imino] methyl]pyridin-2-yl}methyl)amino]-N-[2- (dimethylamino)ethyl]-N- ethylacetamide N 1H-NMR(300 MHz, CD₃OD), δ 8.75 (d, 1H), 7.70 (s, 1H), 7.60 (d, 1H), 7.30 (m,5H), 5.70 (s, 1H), 4.50 (s, 2H), 4.25 (m, 2H), 3.80 (m, 3H), 3.00 (s,6H), 2.60 (m, 4H), 1.20 (t, 3H)

90 2-[({4-[5-benzyl-3- (trifluoroacetyl)-1,3- oxazinan-2-yl]pyridin-2-yl}methyl)amino]-N-[2- (dimethylamino)- ethyl]-N- ethylacetamide Y1H-NMR (300 MHz, CD₃OD), δ 8.70 (d, 1H), 7.40 (m, 2H), 7.25 (m, 5H),4.50 (s, 1H), 4.20 (m, 4H), 3.80 (m, 3H), 3.05 (s, 6H), 2.60 (m, 4H),1.20 (t, 3H).

91 N-[2--(dimethylamino)- ethyl]-N-ethyl-2- [({4-[7-(trifluoroacetyl)-5-oxa-7-azaspiro[2.5]- octan-6-yl]pyridin-2- yl}methyl)amino]- acetamideY 1H-NMR (300 MHz, CD₃OD), δ ppm: 8.8 (d, 1H), 7.5 (m, 2H), 6.7 (m,1H),4.5 (s, 2H),4.2 (s, 2H), 3.7 (t, 2H), 3.4 (m, 6H), 3.0 (m, 8H), 1.2(m, 3H), 0.6 (m, 2H), 0.4 (m, 2H).

92 N-[(2- fluorophenyl)methyl]-2- [({4-[[(2- hydroxyethyl)imino]-methyl]pyridin-2- yl}methyl)amino]-N- methylacetamide O 1H-NMR (300 MHz,CD₃OD), δ 8.55 (d, 1H), 7.55 (s, 1H), 7.50 (s, 1H 7.30 (m, 3H), 7.10 (m,3H), 5.50 (s, 1H), 4.70 (s, 2H),4.50 (s, 2H), 3.30 (s, 2H), 3.00 (m,6H), 2.60 (m, 4H).

93 2-[({2-[({2-[2- (benzyloxy)phenyl]- ethyl}amino)methyl]- pyridin-4-yl}methylidene)- amino]ethan-1-ol N NMR (300 MHz, chloroform-d): δ ppm8.60 (m, 1H), 8.20 (s, 1H), 6.90 (m, 3H), 5.10 (s, 2H), 2.96 (m, 4H).

94 N-(2-cyanoethyl)-N- ethyl-2-[({4-[[(2- hydroxyethyl)imino]-methyl]pyridin-2- yl}methyl)amino]- acetamide O 1H NMR (300 MHz,chloroform-d): δ ppm 8.58 (d, 1H), 8.29 (s, 1H), 7.65 (s, 1H), 7.44 (d,1H), 3.97 (s, 2H), 3.91 (m, 2H), 3.78 (m, 2H), 3.55 (t, 2H), 3.50 (s,2H), 3.36 (q, 2H), 2.68 (m, 4H), 1.17 (t, 3H).

95 (2S)-2-[({4-[[(2- hydroxyethyl)imino]- methyl]pyridin-2-yl}methyl)amino]-4- methyl-1-(piperidin-1- yl)pentan-1-one O 1H NMR (300MHz, chloroform-d): δ ppm: 8.7 (s, 1H), 8.3 (s, 1H), 7.7 (s, 1H), 7.5(d, 1H), 4.1-3.3 (m, 1H), 1.7-1.4 (m, 8H), 0.9 (d, 6H).

96 2-[{4-[([(2- hydroxyethyl)imino]- methyl]pyridin-2-yl}methyl)amino]-N- methyl-N-(2- phenylethyl)acetamide O 1H NMR (300MHz, chloroform-d): δ ppm: 8.6 (d, 1H), 8.3 (s, 1H), 7.6 (d, 1H), 7.4(s, 1H), 7.3-7.1 (m, 5H), 4.0-3.8 (m, 11H), 2.1 (m, 1H), 3.4 (m, 2H),3.0 (s, 2H), 2.9 (m, 5H).

97 2-{[({4- [(dimethylamino)- methyl]cyclohexyl}- methyl)amino]-methyl}pyridine-4- carbaldehyde Z 1H NMR (300 MHz, Methanol-d4), δ ppm:8.89 (d, 1H), 8.26 (s, 1H), 2.91 (s, 6H), 1.95 (m, 6H).

98 2-({[(2E)-4- (dimethylamino)but- 2-en-1-yl]amino}- methyl)pyridine-4-carbaldehyde Z 1H NMR (300 MHz, Methanol-d4), δ ppm: 8.91 (d, 1H),6.25 (m, 2H), 5.75 (s, 1H), 2.93 (s, 6H).

99 2-({[(2Z)-4- (dimethylamino)- but-2-en-1-yl]amino}methyl)- pyridine-4-carbaldehyde AA 1H-NMR (300 MHz, Methanol-d4), δ ppm: 8.8 (d, 1H), 6.2(m, 2H), 4.6 (s, 2H), 2.9 (s, 6H)

100 2-({[(1-methyl- piperidin-4- yl)methyl]amino}- methyl)pyridine-4-carbaldehyde Z 1H NMR (300 MHz, Methanol-d4), δ ppm: 8.97 (d, 1H), 5.79(s, 1H), 2.89 (s, 3H), 1.69 (m, 2H)

101 N-[2- dimethylamino)- ethyl]-N- ethyl-2-{[(4- formylpyridin-2-yl)methyl]amino}- acetamide AA 1H NMR (300 MHz, CD3OD), δ ppm: 8.94 (d,1H), 8.42 (s, 1H), 2.99 (s, 6H), 1.29 (t, 3H).

102 2-[({2-oxo-2- [(2R)-2-(pyrrolidin-1- ylmethyl)pyrrolidin-1-yl]ethyl}amino)- methyl]pyridine-4- carbaldehyde AB 1H NMR (300 MHz,CD3OD), δ ppm: 8.91 (m, 1H), 8.36 (s, 1H), 2.27-2.00 (m, 7H), 1.85 (m,1H)

103 2-({[2-(4- methylpiperazin-1- yl)-2-oxoethyl]- amino}methyl)-pyridine-4- carbaldehyde AB 1H NMR (300 MHz, Methanol-d4), δ ppm: 8.89(d, 1H), 5.74 (s, 1H), 4.51 (m, 4H), 2.97 (s, 3H).

104 N-[(1-ethylpyrrolidin- 2-yl)methyl]-2-{[(4- formylpyridin-2-yl)methyl]amino}- acetamide AB 1H-NMR (300 MHz, MeOD), δ ppm: 8.50 (d,1H), 7.50 (s, 1H), 2.10 (m, 2H), 1.40 (t, 3H)

105 N,N-diethyl-2-{[(4- formylpyridin-2- yl)methyl]amino}- acetamide AB1H-NMR (300 MHz, MeOD), δ ppm: 8.90 (d, 1H), 8.40 (s, 1H), 3.20 (q, 2H),1.20 (t, 3H)

106 2-({[2-(4- benzylpiperidin- 1-yl)-2-oxoethyl]- amino}methyl)-pyridine-4- carbaldehyde AB 1H NMR (300 MHz, Methanol-d4), δ ppm: 8.95(d, 1H), 7.22 (m, 5H), 5.79 (s, 1H), 1.86 (m, 1H).

107 2-({[4-(diethylamino)- butyl]amino}- methyl)pyridine-4- carbaldehydeZ 1H-NMR (300 MHz, MeOD), δ ppm: δ 8.90 (d, 1H), 8.48 (s, 2H), 1.90 (m,4H), 1.40 (t 6H)

108 2-({[4-(dimethylamino)- butyl]amino}methyl)- pyridine-4-carbaldehyde Z 1H-NMR (300 MHz, MeOD), δ ppm: 8.85 (d, 1H), 8.30 (s,1H), 2.90 (s, 6H), 1.90 (m, 4H)

109 2-[({4-[benzyl- (cyclopropyl)amino]- butyl}amino)methyl]-pyridine-4-carbaldehyde AC 1H-NMR (300 MHz, CDCl3), δ ppm: 8.90 (d, 1H),8.40 (s, 1H), 7.50 (m, 5H), 0.90 (m, 4H)

110 2-({[2- (dimethylamino)- ethyl]amino}methyl)- pyridine-4-carbaldehyde Z 1H NMR (300 MHz, Methanol-d4), δ ppm: 8.94 (d, 1H), 5.76(s, 1H), 3.76 (m, 4H), 3.05 (m, 6H)

111 2-({[3-(pyrrolidin-1- yl)propyl]amino}- methyl)pyridine-4-carbaldehyde Z 1H-NMR (300 MHz, Methanol-d4), δ ppm: 8.80 (s, 1H), 5.70(s, 1H), 4.10-3.40 (m, 6H), 2.98-2.40 (m, 2H).

112 N-[4-(diethylamino)- butyl]-2-{[(4- formylpyridin-2-yl)methyl]amino}- acetamide AB 1H NMR (300 MHz, Methanol-d4), δ ppm:8.84 (d, 0.5H), 5.71 (s, 1H), 1.61 (m, 2H), 1.35 (m, 6H).

113 N-(1-benzylpyrrolidin- 3-yl)-2-{[(4- formylpyridin-2-yl)methyl]amino}- acetamide AB 1H-NMR (300 MHz, MeOD), δ ppm: 8.90 (m,2H), 8.30 (m, 1H), 7.40 (m, 5H), 2.00 (m, 2H)

114 2-({[5- (dimethylamino)- pentyl]amino}methyl)- pyridine-4-carbaldehyde Z 1H-NMR (300 MHz, MeOD), δ ppm: 8.80 (d, 1H), 8.20 (s,1H), 2.90 (s, 6H), 1.80 (m, 4H)

115 N-[4- (diethylamino)butyl]- 2,2,2-trifluoro-N-[(4- formylpyridin-2-yl)methyl]acetamide AD 1H NMR(300 MHz, CDCl₃), δ ppm: 10.09 (d, 1H),8.83 (dd, 1H), 1.76 (m, 4H), 1.33 (m, 6H)

116 N-[2- (diethylamino)- ethyl]-N- ethyl-2-{[(4- formylpyridin-2-yl)methyl]amino}- acetamide AA 1H NMR (300 MHz, CD₃OD): δ ppm 8.87 (d,1H), 8.19 (s, 1H), 8.00 (dd, 1H),5.73 (s, 1H), 4.74 (s, 2H), 4.39 (s,2H), 3.03 (t, 2H), 3.49- 3.28 (m, 8H), 1.38 (t, 6H), 1.29 (t, 3H).

117 2-[({[3- (dimethylamino)- cyclopenyl]methyl}- amino)methyl]pyridine-4- carbaldehyde Z 1H-NMR (300 MHz, CDCl₃), δ ppm: 8.90 (s, 1H),5.80 (s, 1H), 2.80 (s, 6H), 2.50-2.00 (m, 6H).

118 N-[2-(dimethylamino)- 2-methylpropyl]- N-ethyl-2-{[(4-formylpyridin-2- yl)methyl]amino}- acetamide AA 1H NMR (300 MHz,DMSO-d₆) δ 10.1 (s, 1H), 8.91 (d, 1H), 4.49 (s, 2H),4.24 (s, 2H), 2.72(s, 6H), 1.34 (s, 6H), 1.12 (t, 3H).

119 N-ethyl-2-{[(4- formylpyridin-2- yl)methyl]amino}- N-[(1-methylpyrrolidin-2- yl)methyl]acetamide AA 1H NMR(300 MHz, CD₃OD): δ ppm8.26 (s, 1H), 4.77 (s, 2H), 4.45 (s, 2H), 3.21 (m, 1H), 3.03 (s, 3H),2.20-2.07 (m, 2H), 1.29 (t, 3H).

120 2-({methyl[2-oxo-2- (piperidin-1- yl)ethyl]amino}-methyl)pyridine-4- carbaldehyde Z 1H-NMR (300 MHz, CDCl₃): δ 10.08 (s,1H), 8.78 (d, 1H), 7.92 (s, 1H), 7.58 (d, 1H).

N-{[2-({[4-(diethylamino)butyl]amino}methylpyridin-4-yl]methyl}-2,2,2-trifluoroacetamide(#1)

Synthetic Route A

General Procedure A (Reductive Amination)

A solution of aldehyde(2,2,2-trifluoro-N-[(2-formylpyridin-4-yl)methyl]acetamide) and amine((4-aminobutyl)diethylamine) (1.3 equiv.) in 1,2-dichloroethane wasstirred for 2 h at room temperature, before NaBH(AcO)₃ (2 eq) was added.The mixture was stirred overnight at room temperature. The solvents wereremoved in vacuo and the residue was purified by preparative TLC (40%MeOH in DCM). The title product was isolated as colorless oil as theacetate salt. ¹H NMR (300 MHz, CD₃OD) δ ppm: 8.57 (d, 1H), 7.38 (s, 1H),7.29 (d, 1H), 4.53 (s, 2H), 4.13 (s, 2H), 3.13 (q, 4H), 3.04 (t, 2H),2.91 (t, 2H), 1.94 (s, 6H), 1.77 (m, 4H), 1.25 (t, 6H). ES-MS: 361[M+H].

[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methanamine (#5)

Synthetic Route B

General Procedure B (Amines from Tert-Butyl Carbamates)

Concentrated hydrochloric acid was added dropwise to the tert-butylcarbamate (tert-butylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-[4-(diethylamino)butyl]carbamate(I)) at 0° C. The resulting solution was reduced to dryness in vacuo toyield the title product as colorless solid as the hydrochloric acidsalt. ¹H-NMR (300 MHz, MeOD): δ 8.82 (d, 1H), 8.05 (s, 1H), 7.80 (d,1H), 4.65 (s, 2H), 4.05 (s, 2H), 3.20 (m, 9H), 1.85 (m, 4H), 1.30 (t,6H) ppm. ES-MS: 265 [M+H+].

N-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-({4-[(trifluoroacetamido)methyl]pyridin-2-yl}methyl)acetamide(#6)

Synthetic Route C

General Procedure C (Formation of Trifluoroacetamide orTrifluoroacetate)

Trifluoroacetic anhydride (2.2 equiv.) was added dropwise to a solutionof the amine([2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methanamine) (1equiv.) and DIPEA (2.5 equiv.) in anhydrous DCM at 0° C. The mixture wasallowed to warm to room temperature and stirred for 12 hours. Quenchedwith sat. NaHCO₃ (aq.). Aqueous work up gave the title compound. ¹H-NMR(300 MHz, CDCl3): δ 11.60, 11.45 (d, 1H), 9.10, 8.70 (d, 1H), 8.45, 8.40(s, 1H), 7.20, 7.10 (d, 1H), 4.70 (d, 2H), 4.50 (t, 2H), 3.10 (m, 4H),1.50 (t, 6H) ppm. ES-MS: 457 [M+1].

[2-({[4-(azetidin-1-yl)butyl]amino}methyl)pyridin-4-yl]methanamine (#7)

Synthetic Route D

General Procedure B from tert-butylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-[4-(azetidin-1-yl)butyl]carbamateyielded the hydrochloric acid salt of the title product as colorlesssolid. ¹H-NMR (300 MHz, CDCl₃): δ 8.70 (d, 1H), 7.60 (s, 1H), 7.40 (d,1H), 4.40 (s, 2H), 4.20 (s, 2H), 3.20 (s, 4H), 2.20 (m, 2H), 1.80 (m,4H), 1.20 (m 4H) ppm. ES-MS: 249 [M+1].

N-{[2-({[4-(dimethylamino)butyl]amino}methyl)pyridin-4-yl]methyl}cyclopropanamine(#10)

Synthetic Route E

By General Procedure A from(4-[(cyclopropylamino)methyl]pyridine-2-carbaldehyde and(4-aminobutyl)dimethylamine) (1.0 equiv.). Purification by columnchromatography (CH2Cl2/MeOH/NH₄OH, 90:10:1) yielded the title compoundas a colorless glue. ¹H NMR (300 MHz, CDCl₃): δ 8.42 (d, 1H), 7.22 (S,1H), 6.95 (m, 1H), 3.70 (s, 2H), 3.65 (s, 2H), 2.65 (m, 2H), 2.25 (m,2H), 2.20 (s, 6H), 2.15 (m, 1H), 2.10-2.00 (m, 4H), 1.62-1.52 (m, 2H),0.5-0.38 (m, 4H).

2-{[(4-{[(cyanomethyl)amino]methyl}pyridin-2-yl)methyl]amino}-N,N-dimethylacetamide(#13)

Synthetic Route F

General Procedure D (Acids from Tert-Butyl Esters or Amines fromTert-Butyl Carbamates)

Trifluoroacetic acid (100 equiv.) was added to a solution of thetert-butyl carbamate (or tert-butyl ester)(tert-butylN-[(4-{[(cyanomethyl)amino]methyl}pyridin-2-yl)methyl]-N-[(dimethylcarbamoyl)methyl]carbamate)(1 equiv.) in DCM at 0° C. The mixture was stirred at room temperaturefor 3 h. Evaporated to dryness to give the title product astrifluoroacetic acid salt. ¹H NMR (300 MHz, methanol-d₄): δ ppm 8.72 (d,1H), 7.57 (s, 1H), 7.53 (d, 1H), 4.37 (s, 2H), 4.36 (s, 2H), 4.29 (s,2H), 4.18 (s, 2H), 3.00 (s, 6H). ES-MS: 262 [M+1].

2-({[4-({[2-(dimethylamino)ethyl]amino}methyl)pyridin-2-yl]methyl}amino)-N,N-dimethylacetamide(#15)

Synthetic Route G

General Procedure E (Hydrolysis of Trifluoroacetamide)

KOH (1.0 M in H₂O, 2.0 equiv.) was added to a solution of thetrifluoroacetamide(N-{[4-({[2-(Dimethylamino)ethyl]amino}methyl)pyridin-2-yl]methyl}-N-[(dimethylcarbamoyl)methyl]-2,2,2-trifluoroacetamide)MeOH/H₂O (1:1 vol). Stirred at 60° C. for about 1.0 h. Evaporated todryness. Aqueous work up gave the title product as oil. ¹H NMR (300 MHz,methanol-d₄): δ ppm 8.44 (d, 1H), 7.47 (s, 1H), 7.32 (d, 1H), 3.90 (s,2H), 3.83 (s, 2H), 3.50 (s, 2H), 2.96 (d, 6H), 2.71 (t, 2H), 2.49 (t,2H), 2.25 (s, 6H). ES-MS: 294 [M+1].

Benzyl(methyl){3-[({4-(methylamino)methylpyridin-2-yl}methyl)amino]propyl}amine (#17)

Synthetic Route H

General Procedure B from tert-butylN-{3-[benzyl(methyl)amino]propyl}-N-({4-[(methylamino)methyl]pyridin-2-yl}methyl)carbamategave the hydrochloric acid salt of title compound. ¹H NMR (300 MHz,CDCl₃), δ ppm: 8.8 (d, 1H), 8.0-7.4 (m, 7H), 4.6-4.2 (m, 6H), 3.9 (s,2H), 3.5-3.2 (m, 4H), 3.0-2.7 (m, 4H), 2.4-2.2 (m, 2H), 1.2-0.9 (m, 4H).

2-[({4-[(cyclopropylamino)methyl]pyridin-2-yl}methyl)amino]-N-{1-[(2-ethoxyphenyl)methyl]-piperidin-4-yl}acetamide(#20)

Synthetic Route I

By General procedure B from tert-butylN-({4-[(cyclopropylamino)methyl]pyridin-2-yl}methyl)-N-[({1-[(2-methoxyphenyl)methyl]piperidin-4-yl}carbamoyl)methyl]carbamateto give the title product as colorless sticky gum. ¹H-NMR (300 MHz,CD₃OD): δ 8.78 (m, 1H), 7.90 (d, 1H), 7.80 (m, 1H), 7.45 (m, 2H), 7.10(m, 1H), 7.02 (m, 1H), 4.58 & 4.50 (2s, 2H, rotamer), 4.40 & 4.30 (2S,2H; rotamer), 4.10 (m, 1H), 3.90 (m, 5H), 3.60-3.65 (m, 2H), 3.20 (m,2H), 2.60 (m, 1H), 2.18 (m, 2H), 1.82 (m, 2H), 0.9-0.8 (m, 4H).

2-[2-({[3-(dimethylamino)propyl]amino}methyl)pyridin-4-yl]-2-(methylamino)acetonitrile(#24)

Synthetic Route 3

By General procedure D from tert-butylN-({4-[cyano(methylamino)methyl]pyridin-2-yl}methyl)-N-[3-(dimethylamino)propyl]carbamate.Evaporation gave the title product as trifluoroacetic acid salt. ¹H NMR(300 MHz, CD₃OD) δ ppm: 8.83 (d, 1H), 7.75 (s, 1H), 7.68 (dd, 1H), 4.54(s, 3H), 3.27 (m, 4H), 2.93 (s, 6H), 2.83 (s, 3H), 2.25 (m, 2H). ES-MS:262 [M+1].

2-[({2-[({4-[benzyl(cyclopropyl)amino]butyl}amino)methyl]pyridin-4-yl}methyl)amino]acetonitrile(#23)

Synthetic Route K

By General procedure D from tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-[(4-{[(cyanomethyl)amino]methyl}pyridin-2-yl)methyl]carbamate.Purification by prep TLC (10% MeOH, 1% NH₄OH in DCM) gave the titlecompound as colorless viscous oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.45 (d,1H), 7.20-7.00 (m, 7H), 4.50 (m, 2H), 3.90 (s, 2H), 3.70 (s, 2H), 3.00(m, 2H), 2.50 (m, 2H), 1.80 (m, 4H), 1.40 (m, 12H) 0.40 (m, 4H), ppm.ES-MS: 378 [M+1].

N-[(2-{[({[2-(Dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2-difluorobutanamide(#32)

Synthetic Route L

By General procedure D from tert-butylN-({4-[(2,2-difluorobutanamido)methyl]pyridin-2-yl}methyl)-N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)carbamateto get the title compound as it's trifluoroacetic acid salt as colorlessoil. ¹H NMR (300 MHz, CD₃OD) δ 8.60 (d, 1H), 7.39 (s, 1H), 7.36 (d, 1H),4.48 (m, 4H), 4.24 (s, 2H), 3.82 (t, 2H), 3.38 (m, 4H), 2.98 (s, 6H),2.32 (m, 2H), 1.24 (t, 3H), 1.02 (t, 3H). ES-MS: 400.61 [M+1]

({[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methyl}carbamoyl)formicacid (#27)

Synthetic Route M

By General Procedure E from tert-butyl({[2-({N-[4-(diethylamino)butyl]-2,2,2-trifluoroacetamido}methyl)pyridin-4-yl]methyl}carbamoyl)formate.Concurrent hydrolysis of the tert-butyl ester and the trifluoroacetamidegave the title product as a yellow sticky gum. ¹H NMR (300 MHz,methanol-d): δ ppm 8.40 (d, 1H), 7.38 (s, 1H), 7.20 (d, 1H), 4.45 (s,2H), 3.80 (s, 2H), 2.70-2.40 (m, 8H), 1.60-1.42 (m, 4H), 1.00 (m, 6H).ES-MS: 337.58 [M+1]

2-({[4-[(N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N,N-dimethylacetamide(#33)

Synthetic Route N

General Procedure D fromtert-butyl-N-({4-[(E)-N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-[(dimethylcarbamoyl)methyl]carbamate)gave the title product as it's trifluoroacetic acid salt as yellow oil.¹H NMR (300 MHz, chloroform-d): δ ppm 8.55 (d, 1H), 8.52 (s, 1H), 7.73(s, 1H), 7.56 (d, 1H), 3.96 (s, 2H), 3.55 (s, 2H), 3.17 (m, 1H), 2.96(d, 6H), 1.00 (m, 4H). ES-MS: 261 [M+1].

N,N-dimethyl-2-[({4-[N-(2-methylcyclopropyl)carboximidoyl]pyridin-2-yl}methyl)amino]acetamide(#35)

Synthetic Route O

General Procedure E fromN-[(dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-({4-[N-(2-methylcyclopropyl)carboximidoyl]pyridin-2-yl}methyl)acetamidegave the title product as yellow oil. ¹H NMR (300 MHz, chloroform-d): δppm 8.57 (d, 1H), 8.36 (s, 1H), 7.64 (s, 1H), 7.42 (d, 1H), 4.99 (s,2H), 3.46 (s, 2H), 2.96 (d, 6H), 2.77 (m, 1H), 1.29 (m, 2H), 1.15 (d,3H), 0.81 (m, 1H). ES-MS: 275 [M+1].

N-{[2-({[2-(ethylsulfanyl)ethyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine(#39)

Synthetic Route P

General Procedure F (Formation of imine)

Amine (cyclopropylamine) (10 equiv.) was added to a solution of aldehyde(2-({[2-(methylsulfanyl)ethyl]amino}methyl)pyridine-4-carbaldehyde) (1equiv.) in DCE. Stirred at room temperature overnight. Evaporated todryness. Purification by preparative TLC (DCM/MeOH/NH₄OH (95/5/1)) gavethe title product as pale yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.5 (d,1H), 8.4 (s, 1H), 7.6 (s, 1H), 7.4 (d, 1H), 3.9 (s, 2H), 3.0 (s, 1H),2.8 (m, 2H), 2.7 (m, 2H) 2.5 (m, 2H), 1.2 (t, 3H), 1.0 (m, 4H).

N-({2-[({3-[benzyl(methyl)amino]propyl}amino)methyl]pyridin-4-yl}methylidene)cyclopropanamine(#41)

Synthetic Route Q

General Procedure D from tert-butylN-{3-[benzyl(methyl)amino]propyl}-N-({4-[(N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)carbamate.Evaporated to dryness to give the title product as trifluoroacetic acidsalt without further purification. ¹H NMR (300 MHz, CDCl₃), δ ppm: 8.55(d, 1H), 8.50 (s, 1H), 7.69 (s, 1H), 7.56 (dd, 1H), 7.28-7.22 (m, 5H),3.89 (s, 2H), 3.50 (s, 2H), 3.20-3.13 (m, 1H), 2.65 (t, 2H), 2.45 (t,2H), 2.11 (s, 3H), 1.82-1.73 (m, 2H), 1.04-0.97 (m, 4H).

N-({2-[({3-[benzyl(methyl)amino]propyl}amino)methyl]pyridin-4-yl}methylidene)cyclopropanamine(#43)

Synthetic Route R

By General Procedure D from tert-butylN-({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-[(2E)-4-(dimethylamino)but-2-en-1-yl]carbamate.Purified by preparative TLC with 1% NH₄OH and 10% MeOH in DCM to givetitle product as light yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm8.59 (d, 1H), 8.41 (s, 1H), 7.55 (s, 1H), 7.42 (d, 1H), 5.71 (m, 2H),3.93 (s, 2H), 3.31 (d, 2H), 3.08 (m, 1H), 2.92 (d, 2H), 2.23 (s, 6H),0.94 (m, 4H). ES-MS: 273 [M+1].

N-{[2-({[4-(azetidin-1-yl)butyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine(#44)

Synthetic Route S

General Procedure D from tert-butylN-[4-(azetidin-1-yl)butyl]-N-({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)carbamate.Evaporation gave the title product as it trifluoroacetic acid saltwithout further purification. ¹H-NMR (300 MHz, CDCl₃): δ 8.50 (d, 1H),8.40 (s, 1H), 7.50 (s, 1H), 7.40 (d, 1H), 3.90 (s, 2H), 3.00 (m, 1H),2.70 (m, 2H), 2.50 (m, 4H), 1.80 (m, 2H), 1.50 (m 6H), 1.00 (m, 4H) ppm.ES-MS: 287 [M+1].

N-{[2-({[4-(dimethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine(#45)

Synthetic Route T

By General Procedure E fromN-({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-[4-(dimethylamino)butyl]-2,2,2-trifluoroacetamide.Evaporated to and the residue was neutralized with cyclopropylamine. 1MKOH solution was added and work-up gave the title product as colorlessviscous oil without further purification. ¹H-NMR (300 MHz, CDCl₃): δ8.50 (s, 1H), 8.30 (s, 1H), 7.50 (s, 1H), 7.30 (d, 1H), 3.90 (s, 2H),2.70 (m, 2H), 2.25 (m, 2H), 2.20 (s, 6H), 1.50 (m, 4H), 1.00 (m, 4H),ppm.

N-{[2-({[5-(dimethylamino)pentyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine(#47)

Synthetic Route U

By General Procedure D from tert-butylN-({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-[5-(dimethylamino)pentyl]carbamate.Evaporated to and the residue was neutralized with cyclopropylamine. 1MKOH solution was added and work-up gave the title product as colorlessviscous oil without further purification. ¹H-NMR (300 MHz, CDCl3): δ8.50 (d, 1H), 8.40 (s, 1H), 7.50 (s, 1H), 7.40 (d, 1H), 3.90 (s, 2H),3.00 (m, 1H), 2.60 (m, 2H), 2.20 (m, 2H), 2.15 (s, 6H), 1.50-1.30 (m,6H), 0.88 (m, 4H) ppm. ES-MS: 289 [M+1].

2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N-[4-(diethylamino)butyl]acetamide(#48)

Synthetic Route V

By General Procedure D fromtert-butyl-N-({4-[(E)-N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-({[4-(diethylamino)butyl]carbamoyl}methyl)carbamate.Purified by preparative TLC (10% MeOH and 1% NH₄OH in DCM) to give titleproduct as yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm 8.61 (d,1H), 8.41 (s, 1H), 7.57 (t, 1H), 7.50 (s, 1H), 7.41 (d, 1H), 3.90 (s,2H), 3.32 (s, 2H), 3.29 (m, 2H), 3.09 (m, 1H), 2.67 (q, 4H), 2.60 (m,2H), 1.56 (m, 4H), 1.09 (t, 6H), 1.03 (m, 4H). ES-MS: 360 [M+1].

2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide(#63)

Synthetic Route X

General Procedure G (Formation of Imine)

To a stirred solution of aldehyde(N-[2-(dimethylamino)ethyl]-N-ethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide)(1 equiv.) in 1,2-DCE and H₂O were added amine (2-cyclohexylethylamine)(1.01 equiv.) and Na₂CO₃ (2 equiv.) at room temperature and stirred for3 hours. Evaporated to dryness. Suspended in DCM, filtered andevaporated to give the title compound as brown oil. ¹H NMR (300 MHz,chloroform-d): δ ppm 8.56 (d, 1H), 8.22 (s, 1H), 7.63 (s, 1H), 7.45 (dd,1H), 3.96 (s, 2H), 3.63 (t, 2H), 3.45 (s, 2H), 3.45-3.34 (m, 2H),3.27-3.19 (m, 2H), 2.43-2.33 (m, 2H), 2.23 and 2.17 (2 singlets, 6H),1.76-1.51 (m, 7H), 1.38-1.25 (m, 1H), 1.22-1.07 (m, 6H), 0.98-0.89 (m,2H). ESI-MS (m/z): 402 [M+1].

2-[({4-[5-benzyl-3-(trifluoroacetyl)-1,3-oxazinan-2-yl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide(#90)

Synthetic Route Y

By General Procedure D from tert-butylN-({4-[5-benzyl-3-(trifluoroacetyl)-1,3-oxazinan-2-yl]pyridin-2-yl}methyl)-N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)carbamatewithout any purification gave the trifluoroacetic acid salt of the titleproduct as yellow oil ¹H-NMR (300 MHz, CD3OD): δ 8.70 (d, 1H), 7.40 (m,2H), 7.25 (m, 5H), 4.50 (s, 1H), 4.20 (m, 4H), 3.80 (m, 3H), 3.05 (s,6H), 2.60 (m, 4H), 1.20 (t, 3H). ES-MS: 536 [M+1]

2-{[({4-[(dimethylamino)methyl]cyclohexyl}methyl)amino]methyl}pyridine-4-carbaldehyde(#97)

Synthetic Route Z

By General Procedure B from tert-butylN-({4-[(dimethylamino)methyl]cyclohexyl}methyl)-N-[(4-formylpyridin-2-yl)methyl]carbamateto yield the title product as colorless solid as the hydrochloric acidsalt. ¹H NMR (300 MHz, methanol-d₄): δ ppm 8.89 (d, 1H), 8.26 (s, 1H),8.05 (d, 1H), 5.73 (s, 1H), 4.66 (s, 2H), 3.12 (d, 2H), 3.04 (d, 2H),2.91 (s, 6H), 1.95 (m, 6H), 1.20 (m, 4H). ES-MS: 290 [M+1].

2-({[(2Z)-4-(dimethylamino)but-2-en-1-yl]amino}methyl)pyridine-4-carbaldehyde(#99)

Synthetic Route AA

General Procedure H (Amines from Tert Butyl Carbamates)

HCl in dioxane (4M) was added to a solution of tert butyl carbamate((Z)-tert-butyl4-(dimethylamino)but-2-enyl((4-formylpyridin-2-yl)methyl)carbamate)) inDCM. The mixture was stirred at room temperature for 1 hour. Evaporatedto give the title compound. 1H NMR (300 MHz, MeOH—d₄: (δ 8.8 (d, 1H),8.0 (s, 1H), 7.7 (d, 1H), 6.2 (m, 2H), 4.5 (m, 2H), 4.1 (m, 2H), 2.9 (s,6H), 2.2 (s, 6H).

2-[({2-Oxo-2-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethyl}amino)methyl]pyridine-4-carbaldehyde(#102)

Synthetic Route AB

By General Procedure B from tert-butylN-[(4-formylpyridin-2-yl)methyl]-N-{2-oxo-2-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethyl}carbamate.Evaporation gave the title product as brown solid. ¹H NMR (300 MHz,CD₃OD), δ ppm: 8.91 (m, 1H), 8.36 (s, 1H), 8.12 (m, 1H), 5.77 (s, 1H),4.85-4.75 (m, 2H), 4.56 (m, 1H), 4.38-4.23 (m, 2H), 4.14 (m, 1H), 3.84(m, 1H), 3.64-3.44 (m, 3H), 3.30-3.19 (m, 2H), 3.11 (m, 1H), 2.27-2.00(m, 7H), 1.85 (m, 1H).

2-[({4-[Benzyl(cyclopropyl)amino]butyl}amino)methyl]pyridine-4-carbaldehyde(#109)

Synthetic Route AC

By General Procedure B from tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-[(4-formylpyridin-2-yl)methyl]carbamate.Evaporation gave the title product as yellow oil. ¹H-NMR (300 MHz,MeOD): δ 8.90 (d, 1H), 8.40 (s, 1H), 8.10 (d, 1H), 7.50 (m, 5H), 5.70(s, 1H), 4.70 (m, 2H), 4.50 (m, 2H), 2.80 (m, 2H), 2.00 (m, 5H), 0.90(m, 4H) ppm.

N-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamide(#115)

Synthetic Route AD

General Procedure I (Trifluoroacetamides from Tert-Butyl Carbamates)

Concentrated H₂SO₄ (2 drops) was added to the tert butyl carbamate(tert-butylN-[4-(diethylamino)butyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate) (1equiv.) in trifluoracetic anhydride at 0° C. The mixture was stirred for2 h at 0° C. Solid NaHCO₃ was added. Diluted with DCM before evaporatingto dryness. Purification by preparative TLC (10% MeOH in DCM) gave thetitle compound as yellow oil. ¹H NMR (300 MHz, CDCl₃), δ ppm: 10.09 (d,1H), 8.83 (dd, 1H), 7.68 (m, 2H), 4.85 (d, 2H), 3.53 (m, 2H), 3.10 (m,6H), 1.76 (m, 4H), 1.33 (m, 6H). ES-MS: 360 [M+1].

Intermediates Ethyl 2-(dimethoxymethyl)pyridine-4-carboxylate(i)—General Procedure 3 (Formation of methyl acetal)

Pyridinium toluene-4-sulphonate (0.1 equiv.) was added to a solution ofaldehyde (ethyl 2-formylpyridine-4-carboxylate) (1.0 equiv.) andtrimethyl orthoformate (6.5 equiv.) in methanol. Heated at 60° C.overnight. Aqueous work up (EtOAc/NaHCO₃) gave the title compound asoil. ¹H NMR (300 MHz, CDCl₃), δ ppm: 8.77 (dd, 1H), 8.09 (s, 1H), 7.82(dd, 1H), 5.44 (s, 1H), 4.41 (q, 2H), 3.42 (s, 6H), 1.41 (t, 3H).

2-(Dimethoxymethyl)pyridine-4-carbaldehyde (ii)—General Procedure K(Reduction of ester to aldehyde)

DIBAL-H (1.5 equiv., 1.0 M in toluene) was added slowly to a solution ofthe ester (Ethyl 2-(dimethoxymethyl)pyridine-4-carboxylate) (1.0 equiv.)in toluene at −78° C. Stirring continued for 1.5 h before the reactionwas quenched by dropwise addition of sat. NH₄Cl. Allowed to warm to roomtemperature. EtOAc and a satd. solution of sodium potassium tartrate(excess) were added and stirring was continued overnight. Aqueous workup gave the title product, which was used without further purification.¹H NMR (300 MHz, CDCl₃), δ ppm: 10.10 (s, 1H), 8.86 (d, 1H), 7.97 (s,1H), 7.68 (dd, 1H), 5.46 (s, 1H), 3.42 (s, 6H).

N-{[2-(dimethoxymethyl)pyridin-4-yl]methylidene}hydroxylamine(iii)—General Procedure L (Formation of Hydroxylamine)

Aldehyde (2-(dimethoxymethyl)pyridine-4-carbaldehyde) (1.0 equiv.) wasdissolved in a mixture of ethanol and water (3:1) and hydroxylaminehydrochloride (1.5 equiv.) was added followed by addition of Na₂CO₃ (1.7equiv.) The suspension was stirred at room temperature for three hoursafter that solvent was removed in vacuum. EtOH was added to the residue,filtered and washed with EtOH. The combined filtrates were evaporated.Triturated with H₂O and filtered to give the title product as colorlesssolid, which was used without further purification. ¹H NMR (300 MHz,CDCl₃), δ ppm: 9.95 (s, 1H), 8.65 (d, 1H), 8.14 (s, 1H), 7.74 (s, 1H),7.50 (d, 1H), 5.45 (s, 1H), 3.43 (s, 6H).

[2-(dimethoxymethyl)pyridin-4-yl]methanamine (iv)—General Procedure M(Hydrogenation to form amines)

A solution of hydroxyl amine(N-{[2-(dimethoxymethyl)pyridin-4-yl]methylidene}hydroxylamine) (1.0equiv.) in MeOH over 10 Pd/C (0.2 equiv. w/w) was charged with H₂ (45psi). The reaction was followed by TLC. Filtered and evaporated to givethe title compound. ¹H NMR (300 MHz, CDCl₃), δ ppm: 8.54 (dd, 1H), 7.50(s, 1H), 7.22 (dt, 1H), 5.35 (s, 1H), 3.91 (s, 2H), 3.40 (s, 6H).

N-{[2-(dimethoxymethyl)pyridin-4-yl]methyl}-2,2,2-trifluoroacetamide (v)

By General Procedure C from 2-(dimethoxymethyl)pyridin-4-yl)methanamine.Evaporated to give the title compound. ¹H-NMR (300 MHz, CDCl3): δ 8.6(d, 1H), 7.4 (s, 1H), 7.2 (d, 1H), 5.4 (s, 2H), 4.6 (d, 2H), 3.4 (s,6H). ES-MS: 277 [M+1] and 321 [M+23]

2,2,2-trifluoro-N-[(2-formylpyridin-4-yl)methyl]acetamide (vi)—GeneralProcedure N (Hydrolysis of acetal)

Concentrated hydrochloric acid (3.5 eq) was added to a solution theacetal(N-{[2-(dimethoxymethyl)pyridin-4-yl]methyl}-2,2,2-trifluoroacetamide)(1 equiv.) in THF. The mixture was stirred for 2 h at 60° C. SolidNaHCO₃ (5 eq) was added at 0° C. and the suspension was filtered toremove the solids, which were washed with dichloromethane. The combinedfiltrates were evaporated to dryness and the residual was purified bycolumn chromatography (0-20% MeOH/DCM) to yield the title product. ¹HNMR (300 MHz, CDCl₃) δ ppm: 10.05 (s, 1H), 8.77 (d, 1H), 7.86 (m, 1H),7.47 (dd, 1H), 7.44 (bs, 1H), 4.65 (d, 2H). ES-MS: 233 [M+H].

[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl][4-(diethylamino)butyl]amine(vii)

By General Procedure A from4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carbaldehyde and4-(diethylamino)butyl]amine. Purification by column chromatography (5%MeOH/DCM) gave the target compound as greenish oil. ¹H-NMR (300 MHz,CDCl₃): δ 8.46 (d, 1H), 7.20 (s, 2H), 7.10 (d, 1H), 4.70 (s, 2H), 3.90(s, 2H), 2.50 (m, 8H), 1.50 (m, 4H), 1.00 (t, 6H, 0.9 (s, 9H, 0.05 (s,6H) ppm.

tert-butylN-[(4-{(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]-N-[4-(diethylamino)butyl]carbamate(viii)—General Procedure O (Boc protection)

Di-tert-butyl dicarbonate (1.2 equiv.) was added to a solution of theamine([(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl][4-(diethylamino)butyl]amine)(1.0 equiv.) and triethylamine (1.3 equiv.) in anhydrous DCM at 0° C.The reaction mixture was stirred at room temperature for 12 hours.Na₂CO₃ was added and the mixture was stirred for 30 min. Evaporated todryness and the residual was extracted with DCM. Evaporation of theextract gave the title compound, which was used without any furtherpurification.

Tert-butylN-[4-(diethylamino)butyl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(ix)—General Procedure P (Removal of Silyl Alcohol Protecting Group)

TBAF (2 equiv.) was added at room temperature to a solution of the silylether (tert-butylN-[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]-N-[4-(diethylamino)butyl]carbamate)(1 equiv.) in THF and the reaction mixture was stirred overnight. Sat.NaHCO₃ (aq) was added. Stirred for 30 min, before work-up with DCM.Purification by column chromatography (DCM, MEOH and NH₄ (aq.)) gave thetitle product as colorless oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.40 (d, 1H),7.00 (m, 2H), 4.60 (s, 2H), 4.40 (m, 2H), 3.20 (m, 2H), 2.40 (m, 6H),1.40 (m, 9H), 1.00 (t, 6H) ppm.

Tert-butylN-[4-(diethylamino)butyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate(x)—General Procedure O (Swern Oxidation)

DMSO (4 equiv.) in anhydrous DCM was cooled to −78° C. and oxalylchloride (2 equiv.) was added drop-by-drop and stirred for 30 min at−78° C. The alcohol (tert-butylN-[4-(diethylamino)butyl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate)(1.0 equiv.) was dissolved in DCM and added slowly at the sametemperature and the reaction mixture was stirred for one hour.Triethylamine (5.0 equiv.) was added and reaction mixture was stirredovernight in the same cooling bath. Quenched with water, and worked upby extraction with DCM. Purification by column chromatography usingethyl acetate/hexane 20-50% yielded the title product. ¹H-NMR (300 MHz,CDCl₃): δ 10.05 (s, 1H), 8.80 (d, 1H), 7.50 (m, 2H), 4.50 (d, 2H), 3.30(d, 2H), 2.50 (m, 6H), 1.4-1.5 (br d, 16H) 1.00 (t, 6H) ppm. ES-MS: 364[M+1].

tert-butylN-[4-(diethylamino)butyl]-N-({4-[(hydroxyimino)methyl]pyridin-2-yl}methyl)carbamate(xi)

By General Procedure L from tert-butylN-[4-(diethylamino)butyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate.Evaporated to dryness. The residue was suspended in dichloromethane,filtered, and the filtrate was evaporated to give the crude titleproduct, which was used without further purification. ¹H-NMR (300 MHz,CDCl₃): δ 8.40 (d, 1H), 8.00 (s, 1H), 7.39 (m, 2H), 4.45 (m, 2H), 3.35(m, 2H), 2.60 (m, 8H), 1.60-1.20 (m, 18H) ppm.

tert-butylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-[4-(diethylamino)butyl]carbamate(xii)

By General Procedure M from tert-butylN-[4-(diethylamino)butyl]-N-({4-[(hydroxyimino)methyl]-pyridin-2-yl}methyl)carbamateto give the title product which was used without further purification.

Ethyl 2-[(4-hydroxybutyl)carbamoyl]pyridine-4-carboxylate (xiii)

By General Procedure A from ethyl 2-formylpyridine-4-carboxylate and4-aminobutan-1-ol. Purification by column chromatography (5% MeOH/DCM)gave the target compound as greenish oil. ¹H-NMR (300 MHz, CDCl₃): δ8.60 (d, 1H), 7.80 (s, 1H), 7.70 (d, 1H), 4.40 (q, 2H), 4.05 (s, 1H),3.60 (m, 2H), 2.80 (m, 2H), 1.70 (m, 4H), 1.40 (t, 3H) ppm.

Ethyl2-({[(tert-butoxy)carbonyl](4-hydroxybutyl)amino}carbonyl)pyridine-4-carboxylate(xiv)—General Procedure R (Boc protection of amine)

Aqueous solution NaHCO₃ (5.0 equiv.) was added to a solution of theamine (ethyl 2-[(4-hydroxybutyl)carbamoyl]pyridine-4-carboxylate) (1.0equiv.) in THF. Stirred for 5 min, before a solution of di-tert-butyldicarbonate (1.2 equiv.) in THF was added. The reaction mixture wasstirred over night at room temperature. Evaporated to dryness andextracted with ethyl acetate to give the title product as a white solid.¹H-NMR (300 MHz, CDCl₃): δ 8.60 (d, 1H), 7.80 (s, 1H), 7.70 (d, 1H),4.50 (m, 2H), 4.40 (q, 2H), 3.60 (m, 2H), 3.30 (m, 2H), 1.70-1.35 (m,17H) ppm.

Ethyl2-{[(4-bromobutyl)[(tert-butoxy)carbonyl]amino]carbonyl}pyridine-4-carboxylate(xv)—General Procedure S (Alcohol to bromide)

CBr₄ (1.1 equiv.) was added to a cold (0° C.) solution of alcohol (ethyl2-({[(tert-butoxy)carbonyl](4-hydroxybutyl)amino}carbonyl)pyridine-4-carboxylate)(1 equiv.) and PPh₃ ((1.1 equivalent). Stirred for 20 min and thenallowed to warm to room temperature over 3-4 hour. Aqueous work up andpurification by column chromatography using (DCM:MeOH (95:5)) gave thetitle product as white solid. ¹H-NMR (300 MHz, CDCl₃): δ 8.50 (d, 1H),7.80 (s, 1H), 7.60 (d, 1H), 4.50 (m, 2H), 4.40 (q, 2H), 3.60 (m, 2H),3.30 (m, 2H), 1.80 (m 2H), 1.60 (m, 2H), 130 (m, 12H) ppm. ES-MS: 415[M+1].

Ethyl2-({[4-(azetidin-1-yl)butyl][(tert-butoxy)carbonyl]amino}carbonyl)pyridine-4-carboxylate(xvi)—General Procedure T (Nucleophilic substitution with amine)

The amine (azetidine hydrochloride) (5.0 equiv.) and subsequently DIPEA(6.0 equiv.) was added to a solution of the bromide (ethyl2-{[(4-bromobutyl)[(tert-butoxy)carbonyl]amino]carbonyl}pyridine-4-carboxylate)(1 equiv.) in acetonitrile. Stirred at 60° C. for 12 hours. Evaporatedto dryness and purified by column chromatography (DCM/MeOH (95:5)) togive the title product as colorless oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.58(d, 1H), 7.70 (s, 1H), 7.60 (d, 1H), 4.40 (m, 2H), 4.30 (q, 2H), 3.60(m, 2H), 3.30 (m, 2H), 3.00 (m, 4H), 1.60 (m 4H), 1.40 (m, 12H) ppm.ES-MS: 392 [M+1].

Tert-butylN-[4-(azetidin-1-yl)butyl]-N-[4-(hydroxymethyl)pyridine-2-carbonyl]carbamate(xvii)—General Procedure U (Reduction of ester to alcohol)

NaBH₄ (2.0 equiv.) was added at room temperature to a solution of ester(ethyl2-({[4-(azetidin-1-yl)butyl][(tert-butoxy)carbonyl]amino}carbonyl)pyridine-4-carboxylate)(1.0 equiv.) in EtOH. Stirred at room temperature for 10 min and thenreflux for 3 hours. Cooled to room temperature and sat. NH₄Cl solutionwas added. Evaporated to dryness and extracted with DCM. Purification bycolumn chromatography (DCM, MeOH and HN₄OH (aq.) (85:10:5) gave thetitle product as viscous oil. 1H-NMR (300 MHz, CDCl₃): δ 8.40 (d, 1H),7.35 (s, 1H), 7.00 (d, 1H), 4.60 (s, 2H), 4.50 (m, 2H), 3.25 (m, 6H),3.20 (m, 2H), 2.00 (m, 2H), 1.40 (m 14H) ppm.

Tert-butylN-[4-(azetidin-1-yl)butyl]-N-(4-formylpyridine-2-carbonyl)carbamate(xviii)—General Procedure V (Dess-Martin oxidation)

1,1,1-Triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (1.1 equiv.) wasadded at 0° C. to a solution of alcohol (tert-butylN-[4-(azetidin-1-yl)butyl]-N-[4-(hydroxymethyl)pyridine-2-carbonyl]carbamate)in anhydrous DCM. Stirred for 10 min. and then allowed to warm to roomtemperature and stirred for two to three hours. KOH solution (1M) wasadded and extraction with DCM gave the title product as light yellowoil. ¹H-NMR (300 MHz, CDCl₃): δ 10.0 (s, 1H), 8.70 (d, 1H), 7.55 (s,1H), 7.50 (d, 1H), 4.50 (m, 2H), 3.20 (m, 2H), 2.50 (m, 4H), 1.80 (m,2H), 1.40 (m 15H) ppm.

Tert-butylN-[4-(azetidin-1-yl)butyl]-N-{4-[((hydroxyimino)methyl]pyridine-2-carbonyl}carbamate(xix)

General Procedure L from tert-butylN-[4-(azetidin-1-yl)butyl]-N-(4-formylpyridine-2-carbonyl)carbamate gavethe title product which was used without further purification. ¹H-NMR(300 MHz, CDCl₃): δ 8.40 (d, 1H), 8.00 (s, 1H), 7.30 (s, 1H), 7.25 (d,1H), 4.40 (m, 2H), 3.20 (m, 4H), 2.00 (m, 2H), 1.60-1.20 (m 20H) ppm.

Tert-butylN-[4-(aminomethyl)pyridine-2-carbonyl]-N-[4-(azetidin-1-yl)butyl]carbamate(xx)

General Procedure M from tert-butylN-[4-(azetidin-1-yl)butyl]-N-{4-[((hydroxyimino)methyl]pyridine-2-carbonyl}carbamategave the title product as colorless oil, which was used without furtherpurification.

N-{[2-(dimethoxymethyl)pyridin-4-yl]methyl}cyclopropanamine (xxi)

By General Procedure A from 2-(dimethoxymethyl)pyridine-4-carbaldehyde,cyclopropylamine, and acetic acid (1 equiv.). Purification by columnchromatography (CH₂Cl2/MeOH, 97:3) gave the title compound as acolorless oil. ¹H NMR (300 MHz, CDCl₃): δ 8.55 (d, 1H), 7.50 (S, 1H),7.22 (m, 1H), 5.40 (s, 1H), 3.90 (s, 2H), 3.40 (s, 6H), 2.18 (m, 1H),0.5 (m, 4H).

4-[(cyclopropylamino)methyl]pyridine-2-carbaldehyde (xxii)

By General Procedure N fromN-{[2-(dimethoxymethyl)pyridin-4-yl]methyl}cyclopropanamine. Usedwithout further purification. ¹H-NMR (300 MHz, CDCl₃): δ 10.02 (s, 1H),8.70 (d, 1H), 7.98 (s, 2H), 7.50 (m, 1H), 3.98 (s, 2H), 2.25 (m, 1H),0.50-0.40 (m, 4H).

Ethyl 2-({[(dimethylcarbamoyl)methyl]amino}methyl)pyridine-4-carboxylate(xxiii)

By General Procedure A from Ethyl 2-formylpyridine-4-carboxylate,N,N-dimethylglycineamide hydrochloride, and triethylamine. Purificationby column chromatography with a gradient of 0-10% MeOH in DCM gave thetitle product as yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm 8.69(s, 1H), 7.94 (s, 1H), 7.72 (d, 1H), 4.40 (q, 2H), 4.03 (s, 2H), 3.48(s, 2H), 2.95 (d, 6H), 1.40 (t, 3H).

2-({[4-(Hydroxymethyl)pyridin-2-yl]methyl}amino)-N,N-dimethylacetamide(xxiv)

By General Procedure U from ethyl2-({[(dimethylcarbamoyl)methyl]amino}methyl)pyridine-4-carboxylate.Purification by column chromatography (10% MeOH and 1% NH₄OH in DCM)gave the title product as light yellow oil. ¹H NMR (300 MHz,chloroform-d): δ ppm 8.44 (d, 1H), 7.47 (s, 1H), 7.30 (d, 1H), 4.67 (s,2H), 3.93 (s, 2H), 3.53 (s, 2H), 2.96 (d, 6H).

Tert-ButylN-[(dimethylcarbamoyl)methyl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(xxv)

General Procedure R from2-({[4-(Hydroxymethyl)pyridin-2-yl]methyl}amino)-N,N-dimethylacetamide.Purification by column chromatography (10% MeOH and 1% NH₄OH in DCM)gave the title product as light yellow oil. ¹H NMR (300 MHz,chloroform-d): δ ppm 8.46 (m, 1H), 7.36 (m, 1H), 7.19 (m, 1H), 4.72 (m,2H), 4.60 (m, 2H), 4.11 (m, 2H), 2.95 (m, 6H), 1.43 (m, 9H).

Tert-ButylN-[(dimethylcarbamoyl)methyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate(xxvi)

General Procedure Q from tert-butylN-[(dimethylcarbamoyl)methyl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.The title product was isolated after extractions as yellow sticky oiland used without further purification. ¹H NMR (300 MHz, chloroform-d): δppm 10.09 (m, 1H), 8.79 (m, 1H), 7.80 (m, 1H), 7.60 (m, 1H), 4.72 (m,2H), 4.18 (m, 2H), 2.97 (m, 6H), 1.43 (m, 9H).

tert-ButylN-[(dimethylcarbamoyl)methyl]-N-({4-[(1E)-(hydroxyimino)methyl]pyridin-2-yl}methyl)carbamate(xxvii)

General Procedure L from tert-ButylN-[(dimethylcarbamoyl)methyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate.The title product was isolated after extraction as light yellow oil andused without further purification. ¹H NMR (300 MHz, methanol-d₄): δ ppm8.47 (m, 1H), 8.10 (d, 1H), 7.61 (d, 1H), 7.46 (m, 1H), 4.58 (m, 2H),4.20 (m, 2H), 2.98 (m, 6H), 1.40 (m, 9H).

Tert-ButylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-[(dimethylcarbamoyl)methyl]carbamate(xxviii)

General Procedure M from tert-ButylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-[(dimethylcarbamoyl)methyl]carbamate.Purification by column chromatography (0-15% MeOH in DCM) gave theproduct as light yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm 8.38(m, 1H), 7.26 (m, 1H), 7.10 (m, 1H), 4.55 (m, 2H), 4.05 (m, 2H), 3.84(m, 2H), 2.89 (m, 6H), 1.36 (m, 9H).

Tert-ButylN-[(4-{[(cyanomethyl)amino]methyl}pyridin-2-yl)methyl]-N-[(dimethylcarbamoyl)methyl]carbamate(xxix)

General Procedure T from tert-butylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-[(dimethylcarbamoyl)methyl]carbamate(1.0 equiv.), DIPEA (2.0 equiv.) and bromoacetonitrile (1.1 equiv.).Purification by preparative TLC (10% MeOH and 1% NH₄OH in DCM) gave thetitle product as light yellow oil. ¹H NMR (300 MHz, methanol-d₄): δ ppm8.42 (d, 1H), 7.46 (s, 1H), 7.33 (s, 1H), 4.58 (m, 2H), 4.19 (m, 2H),3.93 (s, 2H), 3.65 (s, 2H), 2.98 (m, 6H), 1.42 (m, 9H).

2-{[(4-{[(tert-Butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]amino}-N,N-dimethylacetamide(xxx)

By General Procedure A from N,N-dimethylglycineamide hydrochloride,triethylamine, and4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carbaldehyde.Purification by column chromatography (0-10% MeOH in DCM) gave the titleproduct as yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm 8.49 (d,1H), 7.32 (s, 1H), 7.17 (d, 1H), 4.74 (s, 2H), 3.97 (s, 2H), 3.47 (s,2H), 2.95 (d, 6H), 0.95 (s, 9H), 0.11 (s, 6H).

N-[(4-{[(tert-Butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]-N-[(dimethylcarbamoyl)methyl]-2,2,2-trifluoroacetamide(xxxi)

By General Procedure C from2-{[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]amino}-N,N-dimethylacetamide.Evaporation gave the product as yellow oil, which was used withoutfurther purification. ¹H NMR (300 MHz, chloroform-d): δ ppm 8.50 (m,1H), 7.25 (m, 2H), 4.80 (m, 4H), 4.33 (m, 2H), 2.99 (m, 6H), 0.96 (s,9H), 0.13 (s, 6H).

N-[(Dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}acetamide(xxxii)

By General Procedure P fromN-[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]-N-[(dimethylcarbamoyl)methyl]-2,2,2-trifluoroacetamide.Purification by column chromatography (0-10% MeOH in DCM) gave the titleproduct as yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm 8.45 (m,1H), 7.26 (m, 2H), 4.74 (m, 4H), 4.33 (m, 2H), 3.88 (s(br), 1H), 2.92(m, 6H).

N-[(Dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamide(xxxiii)

By General Procedure Q fromN-[(dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-{[4-hydroxymethyl)pyridin-2-yl]methyl}acetamide.Purification by column chromatography (30-60% EtOAc in DCM) gave thetitle product as yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm 10.01(d, 1H), 8.74 (m, 1H), 7.63 (m, 2H), 4.82 (m, 2H), 4.29 (m, 2H), 2.90(m, 6H).

N-[(Dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamide(xxxiv)

By General Procedure A fromN-[(dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamideand N,N-dimethylethylenediamine. Purification by preparative TLC (10%MeOH, 1% NH₄OH in DCM) gave the title product as colorless oil. ¹H NMR(300 MHz, chloroform-d): δ ppm 8.48 (m, 1H), 7.29 (m, 2H), 4.80 (m, 2H),4.37 (m, 2H), 3.84 (m, 2H), 2.98 (m, 6H), 2.71 (t, 2H), 2.49 (t, 2H),2.26 (m, 6H). ES-MS: 390 [M+1].

Ethyl 2-{[(3-hydroxypropyl)amino]methyl}pyridine-4-carboxylate (xxxv)

By General Procedure A from ethyl 2-formylpyridine-4-carboxylate and3-aminopropan-1-ol. Purification by column chromatography (0-5% MeOH inDCM) gave the title compound. ¹H NMR (300 MHz, CDCl₃), δ ppm: 8.72 (d,1H), 7.98 (s, 1H), 7.75 (d, 1H), 4.45 (q, 2H), 3.95 (s, 2H), 3.70 (t,2H), 2.80 (t, 2H), 1.75 (m, 2H), 1.40 (t, 3H).

Ethyl2-[(2,2,2-trifluoro-N-{3-[(trifluoroacetyl)oxy]propyl}acetamido)methyl]pyridine-4-carboxylate(xxxvi)

By General Procedure C from ethyl2-{[(3-hydroxypropyl)amino]methyl}pyridine-4-carboxylate (1 equiv.),DIPEA (7.0 equiv.), and trifluoroacetic anhydride (5.0 equiv.). Thetitle product was isolated after extractions and used without furtherpurification. ¹H-NMR (300 MHz, CDCl₃), (rotamers): δ 8.74 (dd, 0.5H),8.70 (dd, 0.5H), 7.85 (s, 0.5H), 7.83-7.78 (m, 1H), 7.74 (s, 0.5H), 4.81(s, 1H), 4.77 (s, 1H), 4.47-4.35 (m, 4H), 3.70 (t, 1H), 3.58 (t, 1H),2.23-2.13 (m, 1H), 2.10-2.01 (m, 1H), 1.42 (td, 3H).

Ethyl2-{[2,2,2-trifluoro-N-(3-hydroxypropyl)acetamido]methyl}pyridine-4-carboxylate(xxxvii)—General Procedure X (ester hydrolysis)

1M LiOH (aq, 1.0 equiv) was added to a solution of ester (ethyl2-[(2,2,2-trifluoro-N-{3-[(trifluoroacetyl)oxy]propyl}acetamido)methyl]pyridine-4-carboxylate)(1.0 equiv.) in THF-MeOH—H₂O (1:1:1). Stirred at room temperature, whilefollowing the reaction by TLC. Evaporated to dryness and purified byflash chromatography using MeOH:DCM (10:90) to yield the title compound.¹H NMR (300 MHz, CDCl₃), (rotamers) δ ppm: 8.74 (two d, 1H), 7.80 (m,2H), 4.85 (s, 2H), 4.45 (m, 2H), 3.65 (m, 4H), 1.90 (m, 2H), 1.45 (m,3H).

Ethyl2-{[2,2,2-trifluoro-N-(3-oxopropyl)acetamido]methyl}pyridine-4-carboxylate(xxxviii)

By General Procedure Q from ethyl2-{[2,2,2-trifluoro-N-(3-hydroxypropyl)acetamido]methyl}pyridine-4-carboxylateto give the title product. ¹H NMR (300 MHz, CDCl₃), (rotamers) δ ppm:9.80 (two singlets, 1H), 8.70 (two doublets, 1H), 7.80 (m, 2H),4.90/4.75 (two singlets, 2H), 4.45 (m, 2H), 3.95/3.75 (m, 2H), 2.90 (twot, 2H), 1.45 (m, 3H).

Ethyl2-[(N-{3-[benzyl(methyl)amino]propyl}-2,2,2-trifluoroacetamido)methyl]pyridine-4-carboxylate(xxxix)

By General Procedure A from ethyl2-{[2,2,2-trifluoro-N-(3-oxopropyl)acetamido]methyl}pyridine-4-carboxylateand benzyl(methyl)amine to give the title product. ¹H NMR (300 MHz,CDCl3), (rotamers) 8 ppm: 8.7 (dd, 1H), 7.90-7.75 (m, 2H), 7.4-7.2 (m,5H), 4.8 (d, 2H), 4.4 (q, 2H), 4.0 (q, 2H), 3.7-3.4 (m, 2H), 2.8-2.5 (m,2H), 2.0-1.8 (m, 7H), 1.4 (t, 3H).

{2-[({3-[Benzyl(methyl)amino]propyl}amino)methyl]pyridin-4-yl}methanol(xi)

By General Procedure U from ethyl2-[(N-{3-[benzyl(methyl)amino]propyl}-2,2,2-trifluoroacetamido)methyl]pyridine-4-carboxylateusing 5.0 equiv. of NaBH₄. Purification by column chromatography gavethe title product. ¹H NMR (300 MHz, CDCl3) δ ppm: 8.5 (dd, 1H), 7.80 (s,1H), 7.6-7.3 (m, 1H), 4.6 (s, 2H), 4.3 (s, 2H), 3.6-3.2 (m, 4H), 2.8 (s,3H), 2.2-2.0 (m, 2H), 1.7-1.3 (m, 3H).

Tert-butylN-{3-[benzyl(methyl)amino]propyl}-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate

By General Procedure R from,{2-[({3-[benzyl(methyl)amino]propyl}amino)methyl]pyridin-4-yl}methanolto give the title product. ¹H NMR (300 MHz, CDCl3), (rotamers) δ ppm:8.6 (d, 1H), 7.4-7.1 (m, 8H), 4.8 (s, 2H), 4.6-4.4 (m, 2H), 3.4-3.2 (m,2H), 2.5-2.2 (m, 2H), 2.1 (s, 3H), 1.9-1.3 (m, 11H).

Tert-butylN-{3-[benzyl(methyl)amino]propyl}-N-[(4-formylpyridin-2-yl)methyl]carbamate(xlii)

General Procedure Q from tert-butylN-{3-[benzyl(methyl)amino]propyl}-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.¹H NMR (300 MHz, CDCl3), (rotamers) δ ppm: 10.0 (s, 1H), 8.8 (d, 1H),7.7-7.1 (m, 7H), 4.8-4.6 (m, 2H), 3.6-3.2 (m, 4H), 2.5-2.3 (m, 2H), 2.2(s, 3H), 1.9-1.3 (m, 11H).

Tert-butylN-{3-[benzyl(methyl)amino]propyl}-N-({4-[(cyclopropylamino)methyl]pyridin-2-yl}methyl)carbamate(xliii)

By General Procedure A from tert-butylN-{3-[benzyl(methyl)amino]propyl}-N-[(4-formylpyridin-2-yl)methyl]carbamateand cyclopropanamine. Purification by column chromatography usingDCM:MeOH:NH₄OH (8:2:1) gave the title compound. ¹H NMR (300 MHz, CDCl₃),(rotamers) δ ppm: 8.4 (d, 1H), 7.4-7.2 (m, 7H), 4.5 (s, 2H), 3.9 (s,2H), 3.7-3.2 (m, 4H), 2.6-1.7 (m, 8H), 1.4 (d, 9H), 0.6-01.3 (m, 4H).

Ethyl2-({[2-(tert-butoxy)-2-oxoethyl]amino}methyl)pyridine-4-carboxylate(xliv)

Prepared by General Procedure A from ethyl2-formylpyridine-4-carboxylate and tert-butyl 2-aminoacetate. Titlecompound isolated as yellow oil by column chromatography(EtOAc/hexanes). ¹H-NMR (300 MHz, MeOH-d₄): δ 8.7 (d, 1H), 7.8 (s, 1H),7.7 (d, 1H), 4.4 (s, 2H), 4.3 (q, 2), 3.8 (s, 2H), 3.3 (s, 2H), 1.4 (s,(H), 1.3 (t, 3H). ES-MS: 295 [M+1].

Ethyl2-({N-[2-(tert-butoxy)-2-oxoethyl]-2,2,2-trifluoroacetamido}methyl)pyridine-4-carboxylate(xlv)

By General Procedure C from ethyl2-({[2-(tert-butoxy)-2-oxoethyl]amino}methyl)pyridine-4-carboxylate inanhydrous DCM. Aqueous work up gave the title compound, which was usedwithout further purification. ¹H-NMR (300 MHz, CDCl₃), (rotamers): δ 8.7(dd, 1H), 7.8 (ss, 1H), 7.7 (dd, 1H), 4.8 (ss, 2H), 4.3 (q, 2), 4.2 (ss,2H), 1.4 (s, 9H), 1.3 (t, 3H).

2-(N-{[4-(ethoxycarbonyl)pyridin-2-yl]methyl}-2,2,2-trifluoroacetamido)aceticacid (xlvi)

By General Procedure D from ethyl2-({N-[2-(tert-butoxy)-2-oxoethyl]-2,2,2-trifluoroacetamido}methyl)pyridine-4-carboxylate.Evaporation gave the title compound, which was used without furtherpurification. ¹H-NMR (300 MHz, CD₃OD): δ 8.75 (m, 1H), 7.8-8.00 (m, 2H),5.45, 4.99 (2s, 2H; rotamer), 4.20-4.40 (m, 4H), 1.40 (t, 3H).

Ethyl2-({N-[({1-[(tert-butoxy)carbonyl]piperidin-4-yl}carbamoyl)methyl]-2,2,2-trifluoroacetamido}methyl)pyridine-4-carboxylate(xlvii)—General Procedure Y (Amide formation)

An amine (tert-butyl 4-aminopiperidine-1-carboxylate) (2 equiv.) wasadded to a solution of an acid(2-(N-{[4-(ethoxycarbonyl)pyridin-2-yl]methyl}-2,2,2-trifluoroacetamido)aceticacid) (1 equiv.) in DMF.

Cooled to 0° C. before EDC HCl (1.5 equivalent) and ethyl(hydroxyliminocyanoaectate (oxyma; 1.5 equivalent) were added. The reactionmixture was allowed to warm slowly to room temperature and stirredovernight. Aqueous work up and purification by column chromatographygave the title compound as brown foam. ¹H-NMR (300 MHz, CDCl₃): δ 8.70 &8.60 (2d, 1H; rotamer), 7.80 (m, 2H), 4.90 & 4.78 (2s, 2H, rotamer),4.42 (q, 2H), 4.30 & 4.10 (2s, 2H; rotamer), 4.10 (m, 1H), 2.80 (m, 2H)2.0 (m, 2H), 1.48 (s, 9H), 1.40 (t, 3H).

Ethyl2-[(2,2,2-trifluoro-N-{[(piperidin-4-yl)carbamoyl]methyl}acetamido)methyl]pyridine-4-carboxylate(xlviii)

Prepared by General Procedure D from ethyl2-({N-[({1-[(tert-butoxy)carbonyl]piperidin-4-yl}carbamoyl)methyl]-2,2,2-trifluoroacetamido}methyl)pyridine-4-carboxylate.Purification by column chromatography (MeOH/DCM and 1% NH4OH) gave thetitle compound as a brown foam. ¹H-NMR (300 MHz, CD₃OD): δ 8.75 (m, 1H),7.90 (m, 2H), 5.00 & 4.90 (2s, 2H, rotamer), 4.42 (q, 2H), 4.32 & 4.12(2s, 2H; rotamer), 3.95 (m, 1H), 3.40 (m, 2H) 3.10 (m, 2H), 2.10 (m,2H), 1.80 (m, 2H), 1.38 (t, 3H).

Ethyl2-({2,2,2-trifluoro-N-[({1-[(2-methoxyphenyl)methyl]piperidin-4-yl}carbamoyl)methyl]acetamido}methyl)pyridine-4-carboxylate(xlix)

Prepared by General Procedure A from 2-methoxybenzaldehyde and ethyl2-[(2,2,2-trifluoro-N-{[(piperidin-4-yl)carbamoyl]methyl}acetamido)methyl]pyridine-4-carboxylate.¹H-NMR (300 MHz, CDCl₃): δ 8.75 & 8.71 (2d, 1H; rotamer), 7.91-7.78 (m,2H), 7.36 (m, 1H), 7.25 (m, 1H), 7.99-7.86 (m, 2H), 4.95 & 4.72 (2s, 2H,rotamer), 4.45 (q, 2H), 4.30 & 4.08 (2s, 2H; rotamer), 3.83 (m, 4H),3.60 (m, 2H) 2.95 (m, 2H), 2.22 (m, 2H), 1.90 (m, 2H), 1.60 (m, 2H),1.40 (t, 3H).

2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)-N-{1-[(2-methoxyphenyl)methyl]piperidin-4-yl}acetamide(I)

By General Procedure U from ethyl2-({2,2,2-trifluoro-N-[({1-[(2-methoxyphenyl)methyl]piperidin-4-yl}carbamoyl)methyl]acetamido}methyl)pyridine-4-carboxylateusing 5 equiv. of NaBH₄. Purification by column chromatography using 1%MeOH/DCM to 28% MeOH/DCM/1% NH₄OH as elutent to get the product as anoff white solid. ¹H-NMR (300 MHz, CDCl₃): δ 8.42 (d, 1H), 7.42 (d, 1H),7.35 (d, 1H), 7.25 (m, 2H), 7.15 (m, 1H), 6.99-6.85 (m, 2H), 4.65 (s,2H), 3.80 (m, 4H), 3.60 (s, 2H), 3.45 (s, 2H), 3.25 (s, 2H), 2.90 (m,2H), 2.22 (m, 2H), 1.90 (m, 2H), 1.60 (m, 2H).

tert-butylN-{[4-(hydroxymethyl)pyridin-2-yl]methyl}-N-[({1-[(2-methoxyphenyl)methyl]piperidin-4-yl}carbamoyl)methyl]carbamate(li)

By General Procedure R from2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)-N-{1-[(2-methoxyphenyl)methyl]piperidin-4-yl}acetamideto get the title compound as a white foam. ¹H-NMR (300 MHz, CDCl₃): δ9.80 (d, 1H), 8.70 (d, 1H), 7.50-7.40 (m, 2H), 7.25 (m, 2H), 7.10-6.80(m, 2H), 4.70 (s, 2H), 4.50 (d, 2H), 4.12 (d, 2H), 3.98 (m, 3H), 3.85(s, 3H), 3.35 (d, 2H), 2.80-2.50 (m, 2H), 2.10-1.80 (m, 4H), 1.40, 1.20(ss, 9H).

tert-butylN-[(4-formylpyridin-2-yl)methyl]-N-[({1-[(2-methoxyphenyl)methyl]piperidin-4-yl}carbamoyl)methyl]carbamate(lii)

By General Procedure Q from tert-butylN-{[4-(hydroxymethyl)pyridin-2-yl]methyl}-N-[({1-[(2-methoxyphenyl)methyl]piperidin-4-yl}carbamoyl)methyl]carbamateto get the title compound as a light yellow foam. ¹H-NMR (300 MHz,CDCl₃): δ 10.00 (s, 1H), 9.00-8.20 (m, 2H), 7.76 (2d, 1H; rotamer), 7.40(m, 1H), 7.30 (m, 1H), 7.00-6.80 (m, 2H), 4.52 (m, 2H), 4.42 (q, 2H),4.20 & 4.00 (2s, 2H; rotamer), 3.90 (m, 4H), 3.55 (s, 2H) 3.00 (m, 2H),2.22 (m, 2H), 2.00 (m, 2H), 1.80-1.50 (m, 4H), 1.40, 1.20 (2s, 9H;rotamer).

tert-butylN-({4-[(cyclopropylamino)methyl]pyridin-2-yl}methyl)-N-[({1-[(2-methoxyphenyl)methyl]piperidin-4-yl}carbamoyl)methyl]carbamate(liii)

By General Procedure A from tert-butylN-[(4-formylpyridin-2-yl)methyl]-N-[({1-[(2-methoxyphenyl)methyl]piperidin-4-yl}carbamoyl)methyl]carbamateand cyclopropylamine. Purification by column chromatography(CH2Cl2/MeOH/NH₄OH, 90:10:1) gave the title compound as a colorlessglue. ¹H NMR (300 MHz, CDCl₃): δ 9.40 (br s, 1H), 8.62 (2S, 1H;rotamer), 8.40 (s, 1H), 7.50-7.40 (m, 2H), 7.38 (m, 1H), 7.20 (m, 1H),7.00-6.80 (m, 2H), 4.58 & 4.48 (2S, 2H; rotamer), 4.00-3.60 (m, 8H),3.18-2.99 (m, 4H), 2.40-2.20 (m, 2H), 1.80 (m, 2H), 1.70 (m, 2H), 1.40 &1.20 (2S, 9H; rotamer), 1.00 (m, 4H).

Ethyl 2-({[3-(dimethylamino)propyl]amino}methyl)pyridine-4-carboxylate(liv)

By General Procedure A from Ethyl 2-formylpyridine-4-carboxylate and(3-aminopropyl)dimethylamine to get the title compound as dark-orangeoil. ¹H NMR (300 MHz, CDCl₃): δ 8.53 (dd, 1H), 7.70 (s, 1H), 7.56 (dd,1H), 4.24 (q, 2H), 3.86 (s, 2H), 2.60 (t, 2H), 2.26 (t, 2H), 2.10 (s,6H), 1.58 (t, 2H), 1.24 (t, 3H). ES-MS: 266 [M+1].

Ethyl2-({[(tert-butoxy)carbonyl][3-(dimethylamino)propyl]amino}methyl)pyridine-4-carboxylate(lv)

General Procedure R from Ethyl2-({[3-(dimethylamino)propyl]amino}methyl)pyridine-4-carboxylate.

Purification by column chromatography (0-20% MeOH/DCM) gave the titleproduct. ¹H NMR (300 MHz, CDCl₃) δ ppm: 8.68 (dd, 1H), 7.79 (s, 1H),7.73 (d, 1H), 4.62 (d, 2H), 4.41 (q, 2H), 3.34 (m, 2H), 2.26 (m, 2H),2.19 (s, 6H), 1.72 (m, 2H), 1.47 (m, 12H). ES-MS: 366 [M+1].

Tert-butylN-[3-(dimethylamino)propyl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(lvi)

By General Procedure U from Ethyl2-({[(tert-butoxy)carbonyl][3-(dimethylamino)propyl]-amino}methyl)pyridine-4-carboxylate.Purification by column chromatography (0-30% MeOH/DCM) gave the titleproduct. ¹H NMR (300 MHz, CDCl₃) δ ppm: 8.34 (d, 1H), 7.15 (s, 1H), 7.10(d, 1H), 5.18 (bs, 1H), 4.58 (s, 2H), 4.46 (d, 2H), 3.22 (m, 2H), 2.19(m, 2H), 2.10 (s, 6H), 1.61 (m, 2H), 1.36 (d, 9H). ES-MS: 324 [M+1]

Tert-butylN-[3-(dimethylamino)propyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate(lvii)

General Procedure Q from tert-butylN-[3-(dimethylamino)propyl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.Purification by column chromatography (0-20% MeOH/DCM) gave the titleproduct. ¹H NMR (300 MHz, CDCl₃) δ ppm: 10.00 (s, 1H), 8.72 (d, 1H),7.57 (s, 1H), 7.52 (m, 1H), 4.59 (d, 2H), 3.28 (m, 2H), 2.20 (m, 2H),2.12 (s, 6H), 1.66 (m, 2H), 1.38 (d, 9H). ES-MS: 322 [M+1].

Tert-butylN-({4-[cyano(methylamino)methyl]pyridin-2-yl}methyl)-N-[3-(dimethylamino)propyl]carbamate(lviii)—General Procedure Z (Formation of amino alkyl nitriles)

A solution of aldehyde (tert-butylN-[3-(dimethylamino)propyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate)(1 equiv.) and amine (methylamine) (2 equiv.) in anhydrous THF wasstirred overnight at room temperature. Evaporated to dryness andre-dissolved in anhydrous acetonitrile, before TMSCN (1.1 equiv.) wasadded. The mixture was stirred overnight at room temperature. Aqueouswork-up and purification by preparative TLC (3% TEA in 20% MeOH/DCM)gave the title product. ¹H NMR (300 MHz, CDCl₃) δ ppm: 8.58 (d, 1H),7.45 (s, 1H), 7.38 (d, 1H), 4.81 (s, 1H), 4.58 (m, 2H), 3.35 (m, 4H),2.57 (s, 3H), 2.30 (m, 4H), 1.78 (m, 2H), 1.46 (d, 9H). ES-MS: 362[M+1].

Ethyl2-({[(tert-butoxy)carbonyl](4-oxobutyl)amino}methyl)pyridine-4-carboxylate(lix)

By General Procedure Q from ethyl2-({[(tert-butoxy)carbonyl](4-hydroxybutyl)amino}methyl)-pyridine-4-carboxylate.Purified by column chromatography (ethyl acetate/hexane 20-40%) to givethe title product. ¹H-NMR (300 MHz, CDCl₃): δ 9.70 (s, 1H), 8.60 (d,1H), 7.70 (s, 1H), 7.60 (d, 1H), 4.55 (d, 2H), 4.40 (q, 2H), 3.40 (m,2H), 2.480 (m, 2H), 1.85 (m, 2H), 1.50-1.20 (m, 12H) ppm.

Ethyl2-[({4-[benzyl(cyclopropyl)amino]butyl}[(tert-butoxy)carbonyl]amino)methyl]pyridine-4-carboxylate(lx)

By General Procedure A from ethyl2-({[(tert-butoxy)carbonyl](4-oxobutyl)amino}methyl)pyridine-4-carboxylateand N-benzylcyclopropanamine. Purification by column chromatography (5%MeOH/DCM) gave the title compound as colorless oil. ¹H-NMR (300 MHz,CDCl₃): δ 8.60 (d, 1H), 7.70 (s, 1H), 7.60 (d, 1H), 7.10 (m, 5H), 4.50(d, 2H), 4.40 (q, 2H), 3.50 (m, 2H), 3.15 (m, 2H), 2.50 (m, 2H), 1.60(m, 1H), 1.45 (m, 16H), 0.4 (m, 4H) ppm.

Tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(lxi)

By General Procedure U from ethyl2-[({4-[benzyl(cyclopropyl)amino]butyl}[(tert-butoxy)carbonyl]amino)methyl]pyridine-4-carboxylate.Purification by column chromatography (DCM, MeOH and HN₄OH (85; 10:5))gave the title product as viscous oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.45(d, 1H), 7.20 (m, 6H), 7.00 (d, 1H), 4.70 (m, 2H), 4.50 (m, 2H), 3.60(m, 2H), 3.10 (m, 2H), 2.40 (m, 2H), 1.55 (m, 2H), 1.40 (m, 13H) ppm.

Tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-[(4-formylpyridin-2-yl)methyl]carbamate(lxii)

By General Procedure Q from tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.Purification by column chromatography (ethyl acetate/hexane 20-50%) gavethe title product. ¹H-NMR (300 MHz, CDCl₃): δ 10.0 (s, 1H), 8.70 (d,1H), 7.50 (m, 2H), 7.20 (m, 5H), 4.50 (m, 2H), 3.80 (m, 2H), 3.15 (m,2H), 2.50 (m, 2H), 1.60 (m, 2H), 1.45 (m, 12H), 0.40 (m, 4H) ppm.

Tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-({4-[(1E)-(hydroxyimino)methyl]pyridin-2-yl}methyl)carbamate(lxiii)

By General Procedure L from tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-[(4-formylpyridin-2-yl)methyl]carbamate.Evaporation gave the title product, which was used without furtherpurification. ¹H-NMR (300 MHz, CDCl₃): δ 8.45 (d, 1H), 8.00 (s, 1H),7.20-7.00 (m, 7H), 4.50 (m, 2H), 3.70 (s, 2H), 3.20 (m, 2H), 2.50 (m,2H), 1.60-1.20 (m, 14H), 0.40 (m, 4H) ppm.

Tert-butylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-{4-[benzyl(cyclopropyl)amino]butyl}carbamate(lxiv)

By General Procedure M from tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-({4-[(1E)-(hydroxyimino)methyl]pyridin-2-yl}methyl)carbamate.Purification by column chromatography (DCM, MeOH and HN₄OH (85; 10:5))gave the title product as colorless oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.45(d, 1H), 7.20-7.00 (m, 7H), 4.50 (m, 2H), 3.90 (s, 2H), 3.70 (s, 2H),3.00 (m, 2H), 2.50 (m, 2H), 1.80 (m, 4H), 1.40 (m, 12H) 0.40 (m, 4H),ppm.

Tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-[(4-{[(cyanomethyl)amino]methyl}pyridin-2-yl)methyl]carbamate(lxv)

By General Procedure T from tert-butylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-{4-[benzyl(cyclopropyl)amino]butyl}carbamate(1 equiv.) and bromoacetonitrile (1.1 equiv.), using 2 equiv. DIPEA.Aqueous work up gave the title product, which was used without furtherpurification.

Tert-butylN-({[4-(2,2-difluorobutanamido)methyl]pyridin-2-yl}methyl)-N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)carbamate(lxvi)

By General Procedure Y from tert-butylN-{[4-(aminomethyl)pyridin-2-yl]methyl}-N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)carbamate(prepared by synthetic route B (analogously to intermediate xii)) and2,2-difluorobutanoic acid to get the title compound as colorless oil. ¹HNMR (300 MHz, CDCl3) δ 8.48 (m, 1H), 7.27 (m, 1H), 7.07 (m, 2H), 4.55(m, 4H), 4.10 (m, 2H), 3.33 (m, 5H), 2.26 (m, 14H), 1.43 (m, 9H), 1.13(m, 9H).

Tert-butyl({[2-({N-[4-(diethylamino)butyl]-2,2,2-trifluoroacetamido}methyl)pyridin-4-yl]methyl}carbamoyl)formate(lxvii)—General Procedure AA (Amide from acid chloride)

The acid chloride (tert-Butyl 2-chloro-2-oxoacetate (2 equiv.) was addeddropwise to a solution of the amine or trifluoroacetamide(N-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-({4-[(trifluoroacetamido)methyl]pyridin-2-yl}methyl)acetamide)and DIPEA in anhydrous DCM at 0° C. The mixture was allowed to warm toroom temperature and stirred for 12 hours. Quenched with sat. NHaHCO₃(aq.). Aqueous work up (in case of reaction from acetamide: basic workup(NaOH 1N)) product as yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm8.50 (m, 1H), 7.35 (br s, 1H), 7.10-6.90 (m, 2H), 4.70 (m, 2H), 4.50 (m,2H), 3.55-3.30 (m, 2H), 2.58-2.40 (m, 6H), 1.70-1.25 (m, 4H), 1.00 (m,6H). ES-MS: 489.54 [M+1].

tert-ButylN-({4-[(N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-[(dimethylcarbamoyl)methyl]carbamate(lxviii)

By General Procedure F from tert-butylN-[(dimethylcarbamoyl)methyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate(1.0 equiv.) and cyclopropylamine (1.2 equiv.). Evaporated to give thetitle product as yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm 8.55(d, 1H), 8.52 (s, 1H), 7.73 (s, 1H), 7.56 (d, 1H), 3.96 (s, 2H), 3.55(s, 2H), 3.17 (m, 1H), 2.96 (d, 6H), 1.00 (m, 4H). ES-MS: 261 [M+1].

N-[(Dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}acetamide(lxix)

General Procedure C from2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)-N,N-dimethylacetamidegave the title product as yellow oil. ¹H NMR (300 MHz, chloroform-d): δppm 8.45 (m, 1H), 7.26 (m, 2H), 4.74 (m, 4H), 4.33 (m, 2H), 3.88 (s(br),1H), 2.92 (m, 6H).

N-[(Dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamide(lxx)

By General Procedure Q fromN-[(dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}acetamide.Purification by column chromatography (EtOAc 30-60% in DCM) gave thetitle product as yellow sticky oil. ¹H NMR (300 MHz, chloroform-d): δppm 10.01 (d, 1H), 8.74 (m, 1H), 7.63 (m, 2H), 4.82 (m, 2H), 4.29 (m,2H), 2.90 (m, 6H).

N-[(Dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-({4-[(N-(2-methylcyclopropyl)carboximidoyl]pyridin-2-yl}methyl)acetamide(lxxi)

By General Procedure F fromN-[(dimethylcarbamoyl)methyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamide(1.0 equiv.) and 2-methylcyclopropan-1-amine (1.2 equiv.). Evaporationgave the title product as oil. Used with out further purification. ¹HNMR (300 MHz, chloroform-d): δ ppm 8.56 (m, 1H), 8.37 (m, 1H), 7.52 (m,2H), 4.84 (m, 2H), 4.36 (m, 2H), 2.99 (m, 6H), 2.78 (m, 1H), 1.43 (m,1H), 1.25 (m, 1H), 1.15 (m, 3H), 0.83 (m, 1H).

[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl][2-(methylsulfanyl)ethyl]amine(lxxii)

By General Procedure A from4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carbaldehyde and2-(methylsulfanyl)ethan-1-amine. Purification by column chromatography(DCM/MeOH/NH₄OH (90:10:1)) gave the title compound as yellow oil. ¹H NMR(300 MHz, Methanol-d₄): δ 8.4 (d, 1H), 7.3 (s, 1H), 7.1 (d, 1H), 4.7 (s,2H), 3.9 (s, 2H), 2.7 (t, 2H), 2.6 (t, 2H), 2.58 (q, 2H), 1.2 (t, 3H),0.9 (s, 9H).

[2-({[2-(methylsulfanyl)ethyl]amino}methyl)pyridin-4-yl]methanol(lxxiii)

General Procedure P from[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl][2-(methylsulfanyl)ethyl]aminegave the title compound as yellow oil. ¹H NMR (300 MHz, CDCl₃): δ 8.4(d, 1H), 7.3 (s, 1H), 7.1 (d, 1H), 4.7 (s, 2H), 3.8 (s, 2H), 2.7 (t,2H), 2.6 (t, 2H), 2.5 (q, 2H), 1.2 (t, 3H).

2-({[2-(methylsulfanyl)ethyl]amino}methyl)pyridine-4-carbaldehyde(lxxiv)

By General Procedure Q from[2-({[2-(methylsulfanyl)ethyl]amino}methyl)pyridin-4-yl]methanol. Thetitle compound was isolated after evaporation and used without furtherpurification.

tert-ButylN-{3-[benzyl(methyl)amino]propyl}-N-({4-[(N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)carbamate(lxxv)

General Procedure F from tert-butylN-{3-[benzyl(methyl)amino]propyl}-N-[(4-formylpyridin-2-yl)methyl]carbamate(1.0 equiv.) and cyclopropanamine (2 equiv.). Purified by columnchromatography (0-5% MeOH in DCM) to give the title compound.

[(4-{[(tert-Butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl][(2E)-4-(dimethylamino)but-2-en-1-yl]amine(lxxvi)

By General Procedure A from4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carbaldehyde (1.0equiv.), [(2E)-4-aminobut-2-en-1-yl]dimethylamine hydrochloride (1.0equiv.), and triethylamine (1.0 equiv.). Purification by columnchromatography (0-10% MeOH in DCM) gave the title product as yellow oil.¹H NMR (300 MHz, chloroform-d): δ ppm 8.45 (d, 1H), 7.20 (s, 1H), 7.11(d, 1H), 5.67 (m, 2H), 4.68 (s, 2H), 3.85 (s, 2H), 2.89 (s, 2H), 2.19(s, 6H), 0.89 (m, 9H), 0.07 (m, 6H).

tert-ButylN-[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]-N-[(2E)-4-(dimethylamino)but-2-en-1-yl]carbamate(lxxvii)

By General Procedure R from[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl][(2E)-4-(dimethylamino)but-2-en-1-yl]amine.Purification by column chromatography (10% MeOH and 1% NH₄OH in DCM)gave the title product as light yellow oil. ¹H NMR (300 MHz,chloroform-d): δ ppm 8.41 (m, 1H), 7.12 (m, 2H), 5.55 (m, 2H), 4.68 (s,2H), 4.49 (m, 2H), 3.83 (m, 2H), 2.85 (m, 2H), 2.15 (s, 6H), 1.40 (m,9H), 0.91 (s, 9H), 0.07 (s, 6H).

tert-ButylN-[(2E)-4-(dimethylamino)but-2-en-1-yl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(lxxviii)

By General Procedure P from tert-butylN-[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]-N-[(2E)-4-(dimethylamino)but-2-en-1-yl]carbamate.Purification by column chromatography (10% MeOH and 1% NH₄OH in DCM)gave the title product as light yellow oil. ¹H NMR (300 MHz,chloroform-d): δ ppm 8.36 (d, 1H), 7.23 (s, 1H), 7.12 (s, 1H), 5.50 (m,2H), 4.61 (s, 2H), 4.48 (m, 2H), 3.82 (m, 2H), 2.80 (d, 2H), 2.09 (s,6H), 1.39 (m, 9H).

tert-ButylN-[(2E)-4-(dimethylamino)but-2-en-1-yl]-N-[(4-formylpyridin-2-yl)methyl]carbamate(lxxix)

By General Procedure Q from tert-butylN-[(2E)-4-(dimethylamino)but-2-en-1-yl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.Evaporation gave the title product as yellow sticky oil.

Used without further purification. ¹H NMR (300 MHz, chloroform-d): δ ppm10.07 (d, 1H), 8.79 (m, 1H), 7.61 (m, 2H), 5.61 (m, 2H), 4.61 (m, 2H),3.95 (m, 2H), 2.90 (m, 2H), 2.20 (s, 6H), 1.45 (m, 9H).

tert-ButylN-({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-[(2E)-4-(dimethylamino)but-2-en-1-yl]carbamate(lxxx)

By General Procedure F from tert-butylN-[(2E)-4-(dimethylamino)but-2-en-1-yl]-N-[(4-formylpyridin-2-yl)methyl]carbamate(1. Equiv.) and cyclopropanamine (1.2 equiv.). Evaporation gave thetitle product as oil. Used without further purification. ¹H NMR (300MHz, chloroform-d): δ ppm 8.53 (d, 1H), 8.38 (s, 1H), 7.43 (m, 2H), 5.61(m, 2H), 4.51 (m, 2H), 3.89 (m, 2H), 3.06 (m, 1H), 2.94 (m, 2H), 2.24(s, 6H), 1.44 (m, 9H), 0.99 (m, 4H).

tert-ButylN-[4-(azetidin-1-yl)butyl]-N-({4-[(N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)carbamate(lxxxi)

General Procedure F from tert-butylN-[4-(azetidin-1-yl)butyl]-N-(4-formylpyridine-2-carbonyl)carbamate (1.0equiv.) and cyclopropylamine (5 equiv.). Evaporation gave the titleproduct, which was used without further purification. ¹H-NMR (300 MHz,CDCl₃): δ 8.50 (d, 1H), 8.35 (s, 1H), 7.50 (s, 1H), 7.40 (d, 1H), 4.50(m, 2H), 3.20 (m, 2H), 3.00 (m, 2H), 2.90 (m, 2H), 2.10 (m, 2H), 1.70(m, 2H), 1.45 (m, 15H), 1.00 (m, 4H) ppm.

[(4-{[(tert-Butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl][4-(dimethylamino)butyl]amine(lxxxii)

By General Procedure A from4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carbaldehyde and(4-aminobutyl)dimethylamine. Purification by column chromatography(DCM/MeOH (95:5)) gave the title compound as greenish oil. ¹H-NMR (300MHz, CDCl₃): δ 8.48 (d, 1H), 7.25 (s, 2H), 7.10 (d, 1H), 4.70 (s, 2H),3.90 (s, 2H), 2.60 (m, 2H), 2.30 (m, 2H), 2.20 (s, 6H), 1.50 (m, 4H),1.00 (s, 9H, 0.9 (s, 9H), 0.1 (s, 6H) ppm.

N-[(4-{[(tert-Butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]-N-[4-(dimethylamino)butyl]-2,2,2-trifluoroacetamide(lxxxiii)

By General Procedure C from[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl][4-(dimethylamino)butyl]amine.Evaporation gave the title compounds, which was used without furtherpurification.

N-[4-(Dimethylamino)butyl]-2,2,2-trifluoro-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}acetamide(xxxiv)

By General Procedure P fromN-[(4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]-N-[4-(dimethylamino)butyl]-2,2,2-trifluoroacetamide.Purification by column chromatography (DCM/MeOH (90:10)) gave the titlecompound as greenish oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.40 (m, 1H), 7.30(m, 1H), 7.20 (m, 1H), 4.70 (m, 4H), 3.45 (m, 2H), 2.20 (m, 6H), 2.00(s, 2H), 1.50 (m 4H) ppm.

N-[4-(Dimethylamino)butyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamide(lxxxv)

By General Procedure V fromN-[4-(dimethylamino)butyl]-2,2,2-trifluoro-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}acetamide.Aqueous work up gave the title compound as light yellow oil. ¹H-NMR (300MHz, CDCl3): δ 10.0 (s, 1H), 8.80 (d, 1H), 7.55 (s, 2H), 4.80 (m, 2H),3.50 (m, 2H), 2.25 (m, 2H), 2.20 (s, 6H), 1.70 (m, 2H), 1.50 (m, 2H)ppm.

N-({4-[(N-Cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-[4-(dimethylamino)butyl]-2,2,2-trifluoroacetamide(lxxxvi)

By General Procedure F fromN-[4-(dimethylamino)butyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamide(1.0 equiv.) and cyclopropylamine (5 equiv.). Evaporation gave the titleproduct, which was used without further purification. ¹H-NMR (300 MHz,CDCl₃): δ 8.65, 8.50 (d, 1H), 8.45, 8.40 (s, 1H), 7.45 (m, 2H), 4.70 (m,2H), 3.00 (m, 2H), 2.75 (m, 1H), 2.55 (s, 6H), 1.80 (m, 1H), 1.50 (m,5H), 1.00 (m, 4H) ppm.

Ethyl 2-{[(5-hydroxypentyl)amino]methyl}pyridine-4-carboxylate (lxxxvii)

Prepared by General Procedure A from ethyl2-formylpyridine-4-carboxylate (1.0 equiv.) and 5-aminopentan-1-ol (1.2equiv.). Purification by column chromatography (DCM/MeOH (85:15) gavethe title compound as greenish oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.60 (d,1H), 7.80 (s, 1H), 7.00 (d, 1H), 4.30 (q, 2H), 3.90 (s, 2H), 3.50 (t2H), 3.10 (s, 2H), 2.60 (m, 2H), 1.50 (m, 4H), 1.30 (t, 3H) ppm.

Ethyl2-({[(tert-butoxy)carbonyl](5-hydroxypentyl)amino}methyl)pyridine-4-carboxylate(lxxxviii)

Prepared by General Procedure R from ethyl2-{[(5-hydroxypentyl)amino]methyl}pyridine-4-carboxylate. Evaporationgave the title product as white solid. Used without furtherpurification. ¹H-NMR (300 MHz, CDCl₃): δ 8.60 (d, 1H), 7.75 (s, 1H),7.60 (d, 1H), 4.50 (m, 2H), 4.30 (q, 2H), 3.50 (t 2H), 3.20 (m, 2H),2.20 (m, 1H), 1.60-1.20 (m, 18H) ppm.

Ethyl2-({[(tert-butoxy)carbonyl](5-oxopentyl)amino}methyl)pyridine-4-carboxylate(lxxxix)

Prepared by General Procedure Q from Ethyl2-({[(tert-butoxy)carbonyl](5-hydroxypentyl)amino}methyl)pyridine-4-carboxylate.Purification by column chromatography (EtOAc/hexane 20-50%) gave thetitle product. ¹H-NMR (300 MHz, CDCl₃): δ 9.70 (s, 1H), 8.60 (d, 1H),7.75 (s, 1H), 7.60 (d, 1H), 4.50 (m, 2H), 4.40 (q, 2H), 3.20 (m, 2H),2.45 (m, 2H), 1.70-1.30 (m, 17H) ppm.

Ethyl2-({[(tert-butoxy)carbonyl][5-(dimethylamino)pentyl]amino}methyl)pyridine-4-carboxylate(xc)

Prepared by General Procedure A from ethyl2-({[(tert-butoxy)carbonyl](5-oxopentyl)amino}methyl)pyridine-4-carboxylate(1.0 equiv.), dimethylamine hydrochloride (1.2 equiv.), andtriethylamine (1.3 equiv.). Purification by column chromatography(MeOH/DCM (5:95)) gave the title compound as colorless oil. ¹H-NMR (300MHz, CDCl3): δ 8.50 (d, 1H), 7.60 (s, 1H), 7.50 (d, 1H), 4.50 (d, 2H),4.20 (q, 2H), 3.05 (m, 2H), 2.01 (s, 6H), 1.50-1.05 (m, 20H) ppm.

tert-ButylN-[5-(dimethylamino)pentyl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(xci)

Prepared by General Procedure U from ethyl2-({[(tert-butoxy)carbonyl][5-(dimethylamino)pentyl]amino}methyl)pyridine-4-carboxylate.Purification by column chromatography (NH₄OH MeOH/DCM (5:10:85)) gavethe title compound as colorless viscous oil. 1H-NMR (300 MHz, CDCl3): δ8.40 (d, 1H), 7.20 (s, 1H), 7.10 (d, 1H), 4.65 (s, 2H), 4.40 (m, 2H),3.20 (m, 2H), 2.20 (m, 2H), 2.10 (s, 6H), 1.50-1.20 (m, 15H) ppm.

tert-ButylN-[5-(dimethylamino)pentyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate(xcii)

Prepared by General Procedure Q tert-butylN-[5-(dimethylamino)pentyl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.¹H-NMR (300 MHz, CDCl3): δ 10.05 (s, 1H), 8.70 (d, 1H), 7.50 (m, 2H),4.50 (m, 2H), 3.20 (m, 2H), 2.40 (s, 6H), 1.60-1.00 (m, 15H) ppm.

tert-ButylN-({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-[5-(dimethylamino)pentyl]carbamate(xciii)

Prepared by General Procedure F from tert-butylN-[5-(dimethylamino)pentyl]-N-[(4-formylpyridin-2-yl)methyl]carbamate (1equiv.) and cyclopropylamine (5 equiv.). Evaporation gave the titleproduct, which was used without further purification. ¹H-NMR (300 MHz,CDCl₃): δ 8.50 (d, 1H), 8.40 (s, 1H), 7.45 (m, 2H), 4.45 (m, 2H), 3.10(m, 4H), 2.75 (m, 2H), 2.60 (s, 6H), 1.80 (m, 1H), 1.45 (m, 13H), 0.95(m, 4H) ppm.

Ethyl2-{[N-({[4-(diethylamino)butyl]carbamoyl}methyl)-2,2,2-trifluoroacetamido]methyl}pyridine-4-carboxylate(xciv)

By General procedure Y from (4-aminobutyl)diethylamine and2-(N-{[4-(ethoxycarbonyl)pyridin-2-yl]methyl}-2,2,2-trifluoroacetamido)aceticacid. Purification by column chromatography gave the title compound asoil. ¹H NMR (300 MHz, chloroform-d): δ ppm 9.09 (t, 1H), 8.67 (m, 1H),7.83 (m, 2H), 4.85 (m, 2H), 4.42 (q, 2H), 4.17 (m, 2H), 3.26 (m, 2H),2.46 (m, 6H), 1.52 (m, 4H), 1.40 (t, 3H), 0.97 (m, 6H).

N-[4-(Diethylamino)butyl]-2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)acetamide(xcv)

Prepared by General Procedure U from ethyl2-{[N-({[4-(diethylamino)butyl]carbamoyl}methyl)-2,2,2-trifluoroacetamido]methyl}pyridine-4-carboxylate,using 5 equivalents of NaBH₄. Purification by column chromatography with(10% MeOH and 1% NH₄OH in DCM) gave the title product as light yellowoil. ¹H NMR (300 MHz, methanol-d₄): δ ppm 8.45 (d, 1H), 7.47 (s, 1H),7.31 (d, 1H), 4.68 (s, 2H), 3.88 (s, 2H), 3.28 (m, 4H), 2.59 (q, 4H),2.51 (m, 2H), 1.52 (m, 4H), 1.05 (t, 6H).

tert-ButylN-({[4-(diethylamino)butyl]carbamoyl}methyl)-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(xcvi)

Prepared by General Procedure R fromN-[4-(diethylamino)butyl]-2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)acetamide.Purification by column chromatography with (0-20% MeOH in DCM) gave thetitle product as light yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm8.55 (s, 1H), 8.47 (m, 1H), 7.38 (s, 1H), 7.33 (m, 1H), 4.69 (s, 2H),4.61 (m, 2H), 3.99 (m, 2H), 3.58 (t, 0.5H), 3.22 (m, 1H), 2.97 (m, 5H),2.25 (t, 0.5H), 1.82 (m, 1H), 1.64 (m, 4H), 1.36 (m, 9H), 1.22 (m, 6H).

tert-ButylN-({[4-(diethylamino)butyl]carbamoyl}methyl)-N-[(4-formylpyridin-2-yl)methyl]carbamate(xcvii)

Prepared by General Procedure Q from tert-butylN-({[4-(diethylamino)butyl]carbamoyl}methyl)-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.Evaporated to give the title product as a yellow oil, which was usedwithout further purification. ¹H NMR (300 MHz, chloroform-d): δ ppm 11.8(m, 0.5H), 10.1 (s, 1H), 9.17 (m, 0.5H), 8.80 (m, 1H), 7.68 (m, 2H),4.67 (m, 2H), 4.05 (m, 2H), 3.36 (m, 2H), 3.05 (m, 6H), 1.91 (m, 2H),1.66 (m, 2H), 1.38 (m, 9H), 1.17 (m, 6H).

tert-ButylN-({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)-N-({[4-(diethylamino)butyl]carbamoyl}methyl)carbamate(xcviii)

Prepared by General Procedure F from tert-butylN-({[4-(diethylamino)butyl]carbamoyl}methyl)-N-[(4-formylpyridin-2-yl)methyl]carbamateand cyclopropyl amine. Evaporated to give the title product as yellowoil, which was used without further purification. ¹H NMR (300 MHz,chloroform-d): δ ppm 9.60 (t, 1H), 8.51 (m, 1H), 8.39 (s, 1H), 7.46 (m,2H), 4.53 (m, 2H), 3.97 (m, 2H), 3.48 (m, 1H), 3.32 (m, 2H), 2.94 (m,6H), 1.80 (m, 2H), 1.61 (m, 1H), 1.33 (m, 9H), 1.08 (m, 6H).

Ethyl2-{[({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridine-4-carboxylate(xcix)

By General Procedure A from ethyl 2-formylpyridine-4-carboxylate and2-amino-N-[2-(dimethylamino)ethyl]-N-ethylacetamide. ¹H NMR (300 MHz,CDCl₃), δ ppm: 3.47-3.32 (m, 4H), 3.27-3.20 (m, 2H), 2.44-2.35 (m, 2H),2.22 (s, 6H), 1.53 (s, 2H), 1.15-1.06 (m, 3H).

N-[2-(dimethylamino)ethyl]-N-ethyl-2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)acetamide(c)

By General Procedure U from ethyl2-{[({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]-methyl}pyridine-4-carboxylate.¹H NMR (300 MHz, CDCl₃), δ ppm: 8.45 (d, 1H), 7.39 and 7.36 (2 singlets,1H), 7.14 (d, 1H), 4.67 (s, 2H), 3.94 (s, 2H), 3.54-3.47 (m, 4H),3.32-3.24 (m, 2H), 2.60-2.40 (m, 2H), 2.37 and 2.24 (2 singlets, 6H),1.18-1.10 (m, 3H).

tert-ButylN-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(ci)

By General Procedure R fromN-[2-(dimethylamino)ethyl]-N-ethyl-2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)acetamide.¹H NMR (300 MHz, CDCl₃), δ ppm: 8.49 (d, 1H), 7.37 and 7.32 (2 singlets,1H), 7.19 (d, 1H), 4.73 (s, 2H), 4.67 and 4.63 (2 singlets, 2H), 4.20and 4.07 (2 singlets, 2H), 3.48-3.22 (m, 4H), 2.49-2.39 (m, 2H), 2.27and 2.23 (2 singlets, 6H), 1.46 and 1.41 (2 singlets, 9H), 1.21-1.09 (m,3H).

tert-butylN-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)-N-[(4-formylpyridin-2-yl)methyl]carbamate(cii)

By General Procedure Q fromtert-buty-N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.¹H NMR (300 MHz, CDCl₃), δ ppm: 10.07 and 10.06 (2 singlets, 1H), 8.78and 8.75 (2 doublets, 1H), 7.80 and 7.74 (2 doublets, 1H), 7.58 and 7.57(2 singlets, 1H), 4.75 and 4.70 (2 singlets, 2H), 4.24 and 4.09 (2singlets, 2H), 3.54-3.21 (m, 4H), 2.60-2.39 (m, 2H), 2.37, 2.33 and 2.22(3 singlets, 6H), 1.44 and 1.36 (2 singlets, 9H), 1.23-1.08 (m, 3H).

N-[2-(dimethylamino)ethyl]-N-ethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide(ciii)

By General Procedure D fromtert-butyl-N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)-N-[(4-formylpyridin-2-yl)methyl]carbamate.¹H NMR (300 MHz, CD₃OD), δ ppm: 8.94 (d, 1H), 8.42 (s, 1H), 8.17 (dd,1H), 5.79 (s, 1H), 4.85 (s, 2H), 4.44 (s, 2H), 3.86 (t, 2H), 3.49-3.41(m, 4H), 2.99 (s, 6H), 1.29 (t, 3H).

Tert-butylN-({4-[5-benzyl-3-(trifluoroacetyl)-1,3-oxazinan-2-yl]pyridin-2-yl}methyl)-N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)carbamate(civ)—General Procedure AB (Preparation of N-acyl-1,3-oxazinanes)

Optionally substituted 3-aminopropanol (3-amino-2-phenylpropan-1-ol)(1.1 eq) was added to a stirred solution of aldehyde (tert-butyl N-({[2(dimethylamino)ethyl](ethyl)carbamoyl}methyl)-N-[(4-formylpyridin-2-yl)methyl]carbamate)(1.0 eq) in toluene. Stirred at room temperature for 2 h. Anhydride(trifluoroacetic anhydride) (1.5 equiv.) was added dropwise to thesolution followed by 5 equivalent of DIPEA. The mixture was heated at80° C. for two hours. Aqueous work up and purification by HPLC (0.1% TFAsolution/MeOH) gave the title product. ¹H-NMR (300 MHz, CD₃OD): δ 8.50(d, 1H), 7.45 (m, 1H), 7.25 (m, 6H), 4.60 (s, 2H), 4.30 (m, 2H), 3.70(m, 3H), 2.90 (s, 6H), 1.50 (s, 9H), 1.30 (t, 3H) ppm. ES-MS: 636 [M+1]

Ethyl2-{[({4-[(dimethylamino)methyl]cyclohexyl}methyl)amino]methyl}pyridine-4-carboxylate(cv)

By General Procedure A from ethyl 2-formylpyridine-4-carboxylate and{4-[(dimethylamino)methyl]cyclohexyl}methanamine. Aqueous work up gavethe title compound as a yellow oil. Used without further purification.¹H NMR (300 MHz, chloroform-d): δ ppm 8.41 (d, 1H), 8.04 (s, 1H), 7.49(d, 1H), 4.04 (q, 2H), 3.39 (s, 2H), 2.46 (m, 4H), 2.38 (d, 6H), 1.60(m, 4H), 1.34 (t, 3H), 1.03 (m, 2H), 0.68 (m, 4H).

Ethyl2-({[(tert-butoxy)carbonyl]({4-[(dimethylamino)methyl]cyclohexyl}methyl)amino}-methyl)pyridine-4-carboxylate(cvi)

By General Procedure R from ethyl2-{[({4-[(dimethylamino)methyl]cyclohexyl}amino]methyl}pyridine-4-carboxylate.Purification by column chromatography (0-10% MeOH in DCM) gave the titleproduct as light yellow oil. ¹H NMR (300 MHz, chloroform-d): δ ppm 8.62(m, 1H), 7.71 (m, 2H), 4.56 (m, 2H), 4.36 (m, 2H), 3.11 (m, 2H), 2.17(s, 6H), 2.05 (m, 2H), 1.74 (m, 4H), 1.40 (m, 14H), 0.85 (m, 4H).

tert-ButylN-({4-[(dimethylamino)methyl]cyclohexyl}methyl)-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(cvii)

By General Procedure U from ethyl2-({[(tert-butoxy)carbonyl]({4-[(dimethylamino)methyl]cyclohexyl}amino}methyl)pyridine-4-carboxylate)(1.0 equiv.) in EtOH. Stirred at reflux for 2 hours. Cooled to roomtemperature and sat. NH₄Cl solution was added. Evaporated to dryness.Purification by column chromatography (DCM, MeOH (10%) and HN₄OH (1%))gave the title product as light yellow oil. ¹H NMR (300 MHz,chloroform-d): δ ppm 8.41 (d, 1H), 7.17 (m, 2H), 4.65 (s, 2H), 4.50 (d,2H), 3.10 (m, 2H), 2.17 (s, 6H), 2.07 (d, 2H), 1.74 (m, 4H), 1.41 (m,11H), 0.87 (m, 4H).

tert-ButylN-({4-[(dimethylamino)methyl]cyclohexyl}methyl)-N-[(4-formylpyridin-2-yl)methyl]carbamate(cviii)

General Procedure Q from tert-butylN-({4-[(dimethylamino)methyl]-cyclohexyl}methyl)-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamategave the title product as yellow sticky oil without furtherpurification. ¹H NMR (300 MHz, chloroform-d): δ ppm 10.07 (s, 1H), 8.79(m, 1H), 7.61 (m, 2H), 4.63 (d, 2H), 3.18 (m, 2H), 2.22 (s, 6H), 2.10(d, 2H), 1.79 (m, 4H), 1.45 (m, 11H), 0.91 (m, 4H).

Ethyl2-({[(2Z)-4-(dimethylamino)but-2-en-1-yl]amino}methyl)pyridine-4-carboxylate(cix)

By General Procedure A from ethyl 2-formylpyridine-4-carboxylate,(Z)—N1,N1-dimethylbut-2-ene-1,4-diamine. Purification by columnchromatography (DCM/MeOH/NH₄OH, 90:10:1) gave the title compound asyellow viscous oil. ¹H NMR (300 MHz, CDCl₃): δ 8.8 (d, 1H), 7.8 (s, 1H),7.7 (d, 1H), 5.7 (m, 2H), 4.34 (q, 2H), 4.0 (s, 2H), 3.4 (d, 2H), 2.9(d, 2H), 2.2 (s, 6H), 1.32 (t, 3H).

[2-({[(2Z)-4-(Dimethylamino)but-2-en-1-yl]amino}methyl)pyridin-4-yl]methanol(cx)

By General Procedure U from ethyl2-({[(2Z)-4-(dimethylamino)but-2-en-1-yl]amino}methyl)pyridine-4-carboxylate.Purification by column chromatography (DCM/MeOH/NH₄OH, 85:15:1 gave thetitle compound as yellow viscous oil. ¹H NMR (300 MHz, CDCl₃): δ 8.5 (d,1H), 7.2 (s, 1H), 7.1 (d, 1H), 5.8 (m, 2H), 5.4 (s, 2H), 3.9 (s, 2H),3.4 (d, 2H), 2.9 (d, 2H), 2.2 (s, 6H).

tert-ButylN-[(2Z)-4-(dimethylamino)but-2-en-1-yl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(cxi)

By General Procedure R from[2-({[(2Z)-4-(dimethylamino)but-2-en-1-yl]amino}methyl)pyridin-4-yl]methanol.Evaporation gave the title compound as a colorless glue, which was usedwithout further purification. ¹H NMR (300 MHz, CDCl₃): δ 8.5 (d, 1H),7.2 (s, 1H), 7.1 (d, 1H), 5.8 (m, 2H), 4.6 (s, 2H), 4.4 (s, 2H), 3.9 (s,2H), 2.9 (d, 2H), 2.2 (s, 6H), 1.3 (s, 9H).

tert-ButylN-[(2Z)-4-(dimethylamino)but-2-en-1-yl]-N-[(4-formylpyridin-2-yl)methyl]carbamate(cxii)

By General Procedure Q fromtert-butyl-N-[(2Z)-4-(dimethylamino)but-2-en-1-yl]-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.¹H NMR (300 MHz, CDCl₃), (rotamers): δ 10.1 (s, 1H), 8.8 (d, 1H), 7.5(m, 2H), 5.8 (m, 2H), 4.6 (s, 2H), 3.9 (s, 2H), 2.9 (d, 2H), 2.2 (s,6H), 1.3 (s, 9H).

Ethyl2-[(2,2,2-trifluoro-N-{2-oxo-2-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethyl}acetamido)-methyl]pyridine-4-carboxylate(cxiii)

General Procedure Y from (2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidine and2-(N-{[4-(ethoxycarbonyl)pyridin-2-yl]methyl}-2,2,2-trifluoroacetamido)aceticacid gave the title product as yellow oil, which was used withoutfurther purification.

2-({[4-(Hvdroxymethyl)pyridin-2-yl]methyl}amino)-1-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethan-1-one(cxiv)

Prepared by General Procedure U from ethyl2-[(2,2,2-trifluoro-N-{2-oxo-2-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethyl}acetamido)methyl]pyridine-4-carboxylate,using 5 equiv. of NaBH₄. The residue was purified by columnchromatography on silica gel using NH₄OH (2%)+MeOH (20%) in CH₂Cl₂ togive the title as yellow oil. ¹H NMR (300 MHz, CD₃OD), δ ppm: 8.44 (d,1H), 7.49 (s, 1H), 7.31 (d, 1H), 4.90 (s, 2H), 4.69 (s, 2H), 4.24 (m,1H), 3.93 (s, 2H), 3.50-3.36 (m, 3H), 2.67-2.42 (m, 6H), 2.06-1.89 (m,4H), 1.84-1.72 (s, 4H). ESI-MS (m/z): 333 [M+1].

tert-ButylN-{[4-(hydroxymethyl)pyridin-2-yl]methyl}-N-{2-oxo-2-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethyl}carbamate(cxv)

Prepared by General Procedure R from2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)-1-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethan-1-one.Purified by column chromatography on silica gel using NH₄OH (1.5%)+MeOH(15%) in CH₂Cl₂ to give the title compound as brown oil. ¹H NMR (300MHz, CDCl₃), δ ppm: 8.44 (d, 1H), 7.35 and 7.31 (2 singlets, 1H), 7.18(d, 1H), 4.71 and 4.68 (2 singlets, 2H), 4.61 (s, 2H), 4.29 (m, 1H),4.08 and 4.07 (2 singlets, 2H), 3.95 (m, 1H), 3.48-3.01 (m, 7H), 2.87(m, 1H), 2.35 (m, 1H), 2.07-1.93 (m, 7H), 1.43 and 1.41 (2 singlets,9H). ESI-MS: 433 [M+1].

tert-ButylN-[(4-formylpyridin-2-yl)methyl]-N-{2-oxo-2-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethyl}carbamate(cxvi)

General Procedure V from2-({[4-(hydroxymethyl)pyridin-2-yl]methyl}amino)-1-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethan-1-onegave the title compound as yellow oil. ¹H NMR (300 MHz, CDCl₃), δ ppm:10.09 (s, 1H), 8.78 (m, 1H), 7.81 (m, 1H), 7.60 (m, 1H), 4.87-4.51 (m,2H), 4.42-3.87 (m, 2H), 3.66-3.17 (m, 3H), 2.80-2.38 (m, 6H), 2.17-1.74(m, 8H), 1.46 and 1.39 (2 singlets, 9H).

Ethyl2-({[(tert-butoxy)carbonyl](4-oxobutyl)amino}methyl)pyridine-4-carboxylate(cxvii)

By General Procedure Q from ethyl2-({[(tert-butoxy)carbonyl](4-hydroxybutyl)amino}methyl)-pyridine-4-carboxylate.Purified by column chromatography (ethyl acetate/hexane 20-40%) to givethe title product. ¹H-NMR (300 MHz, CDCl₃): δ 9.70 (s, 1H), 8.60 (d,1H), 7.70 (s, 1H), 7.60 (d, 1H), 4.55 (d, 2H), 4.40 (q, 2H), 3.40 (m,2H), 2.480 (m, 2H), 1.85 (m, 2H), 1.50-1.20 (m, 12H) ppm.

Ethyl2-[({4-[benzyl(cyclopropyl)amino]butyl}[(tert-butoxy)carbonyl]amino)methyl]pyridine-4-carboxylate(cxviii)

By General Procedure A from ethyl2-({[(tert-butoxy)carbonyl](4-oxobutyl)amino}methyl)pyridine-4-carboxylateand N-benzylcyclopropanamine. Purification by column chromatography (5%MeOH/DCM) gave the title compound as colorless oil. ¹H-NMR (300 MHz,CDCl₃): δ 8.60 (d, 1H), 7.70 (s, 1H), 7.60 (d, 1H), 7.10 (m, 5H), 4.50(d, 2H), 4.40 (q, 2H), 3.50 (m, 2H), 3.15 (m, 2H), 2.50 (m, 2H), 1.60(m, 1H), 1.45 (m, 16H), 0.4 (m, 4H) ppm.

tert-ButylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate(cxix)

By General Procedure U from ethyl2-[({4-[benzyl(cyclopropyl)amino]butyl}[(tert-butoxy)carbonyl]amino)methyl]pyridine-4-carboxylate.Purification by column chromatography (DCM, MeOH and HN₄OH (85; 10:5))gave the title product as viscous oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.45(d, 1H), 7.20 (m, 6H), 7.00 (d, 1H), 4.70 (m, 2H), 4.50 (m, 2H), 3.60(m, 2H), 3.10 (m, 2H), 2.40 (m, 2H), 1.55 (m, 2H), 1.40 (m, 13H) ppm.

tert-ButylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-[(4-formylpyridin-2-yl)methyl]carbamate(cxx)

By General Procedure Q from tert-butylN-{4-[benzyl(cyclopropyl)amino]butyl}-N-{[4-(hydroxymethyl)pyridin-2-yl]methyl}carbamate.Purification by column chromatography (ethyl acetate/hexane 20-50%) gavethe title product. ¹H-NMR (300 MHz, CDCl₃): δ 10.0 (s, 1H), 8.70 (d,1H), 7.50 (m, 2H), 7.20 (m, 5H), 4.50 (m, 2H), 3.80 (m, 2H), 3.15 (m,2H), 2.50 (m, 2H), 1.60 (m, 2H), 1.45 (m, 12H), 0.40 (m, 4H) ppm.

Reagents Methyl4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carboxylate—GeneralProcedure AC (Formation of Silyl Ether)

Tert-butyldimethylsilyl chloride (1.2 equiv) was added to a solution ofalcohol (4-(hydroxymethyl)pyridine-2-carboxylate) (1. equiv.),triethylamine (2.30 equiv.) and 4-dimethylaminopyridine (0.10 equiv.) indichloromethane at 0° C., Stirred at room temperature overnight. Aqueouswork up and purification by flash chromatography (Hexane-EtOAc, 5-25%)gave the title compound as colorless oil. ¹H-NMR (300 MHz, CD₃OD): δ8.60 (d, 1H), 8.15 (s, 1H), 7.60 (d, 1H), 4.88 (s, 2H), 3.97 (s, 3H),0.98 (s, 9H), 0.15 (s, 6H).

4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carbaldehyde

By General Procedure K from methyl4-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carboxylate (1equiv.). Aqueous work up gave the title compound, which was used withoutfurther purification. ¹H-NMR (300 MHz, CDCl₃): δ 10.11 (s, 1H), 8.75 (d,1H), 7.92 (s, 1H), 7.54 (d, 1H), 7.27 (s, 1H), 4.83 (s, 2H), 0.98 (s,9H), 0.15 (s, 6H).

Ethyl 2-[(chlorocarbonyl)oxy]benzoate—General Procedure AD (acidchloride from carboxylic acid)

The carboxylic acid (Ethyl 2-hydroxybenzoate) (1 eq) in toluene wascooled to 0° C., N, N-dimethyl amine (1 eq) was added, phosgene (1 eq)was added dropwise and stirred at same temperature for 2 h.

The solid was filtered off and the filtrate was concentrated and used asreagent without further purification.

2-amino-(N-[2-(dimethylamino)ethyl]-N-ethyl)acetamide

By General Procedure Y from 2-{[(tert-butoxy)carbonyl]amino}acetic acidand [2-(dimethylamino)ethyl](ethyl)amine. The product was treated withconcentrated hydrochloric acid to get the title compound as hydrochloricacid salt. ¹H NMR (300 MHz, CDCl₃), δ ppm: 3.47-3.32 (m, 4H), 3.27-3.20(m, 2H), 2.44-2.35 (m, 2H), 2.22 (s, 6H), 1.53 (s, 2H), 1.15-1.06 (m,3H).

Tert-butyl 2-[(1EH)-(ethylimino)methyl]pyrrolidine-1-carboxylate

By General Procedure G from tert-Butyl 2-formylpyrrolidine-1-carboxylateand ethylamine. Used without further purification

Ethyl[(1-methylpyrrolidin-2-yl)methyl]amine—General Procedure AE (Aminesfrom amides)

LAH was added to a solution of tert-butyl2-[(1E)-(ethylimino)methyl]pyrrolidine-1-carboxylate in THF and refluxedfor 6 hr. Aqueous work up. NaBH₄ and AcOH were added to a methanolicsolution of the resulting intermediate. Aqueous work up gave the titlecompound as yellow oil. ¹H NMR (300 MHz, CDCl3): δ ppm 3.05 (m, 1H),2.77 (dd, 1H), 2.68 (q, 2H), 2.55 (m, 1H), 2.35 (s, 3H), 2.26-2.15 (m,2H), 1.94 (m, 1H), 1.81-1.57 (m, 5H), 1.12 (t, 3H).

syn-2-(dimethylamino)-N-ethylcyclopentane-1-carboxamide

General Procedure Y from Ethyl[(1-methylpyrrolidin-2-yl)methyl]amine andethylamine gave the product as yellow oil. ¹H NMR (300 MHz, CDCl3): δppm 3.10 (q, 2H), 3.07-2.98 (m, 2H), 2.75 (s, 6H), 2.10-1.81 (m, 6H),1.10 (t, 3H). ESI-MS (m/z): 185 [M+1].

syn-2-[(ethylamino)methyl]-N,N-dimethylcyclopentan-1-amine

General Procedure AE fromsyn-2-(dimethylamino)-N-ethylcyclopentane-1-carboxamide gave the titlecompound as yellow oil ¹H NMR (300 MHz, CDCl3): δ ppm 2.76 (dd, 1H),2.71-2.57 (m, 2H), 2.34 (m, 1H), 2.23 (m, 1H), 2.22 (s, 6H), 2.16 (m,1H), 1.84-1.69 (m, 2H), 1.68-1.56 (m, 3H), 1.51-1.42 (m, 1H), 1.11 (t,3H).

[2-(Dimethylamino)-2-methylpropyl](ethyl)amine

Prepared by General Procedure A from 2-(dimethylamino)-2-methylpropanaland ethylamine to get the title compound as colorless oil. ¹H NMR (300MHz, CDCl₃) δ 2.63 (q, 2H), 2.50 (s, 2H), 2.19 (s, 6H), 1.11 (t, 3H),1.01 (s, 6H).

3-(aminomethyl)-N,N-dimethylcyclopentan-1-amine

General procedure M from 3-(dimethylamino)cyclopentane-1-carbonitrile toget the title product. ¹H-NMR (300 MHz, CDCl₃), δ ppm: 2.8 (m, 2H), 2.5(m, 1H), 2.6 (m, 1H), 2.2 (s, 3H), 2.1 (s, 3H).

Example 2 Histone Lysine Demethylase AlphaLISA Assays for IC50 ValueDetermination

This example demonstrates the ability of compounds of the invention toinhibit the activity in vitro of tested enzymes.

Assays are performed analogously to the protocol described byPerkinElmer (Roy et al. PerkinElmer Technical Note: AlphaLISA #12, April2011)

General Method

Enzymes are dissolved in enzyme buffer and incubated for 10 min beforebeing added to 3% DMSO solutions of compounds in enzyme buffer.Incubated for another 10 minutes, before substrate solution is added andthe reaction mixture is incubated at room temperature. 10 μL acceptorbeads, suspended in Epigenetic Buffer (Perkin Elmer AL008) from stock,are added and the suspension is incubated in the dark at roomtemperature, before a suspension of streptavidin donor beads (PerkinElmer 6760002) in Epigenetic Buffer is added. After incubation at roomtemperature in the dark the plates are read.

Enzymes:

Expression Protein name Vendor/source Sequence organism KDM2B (FBXL10)BPS, Bioscience,   1-650 Bac US KDM3B (JMJD1B) BRIC  842-1761 Bac KDM4A(JMJD2A) BPS, Bioscience,   1-350 E. coli US KDM4B (JMJD2B) BPS   2-500Bac KDM4C (JMJD2C) BRIC, Denmark   1-349 E. coli KDM5C (JARID1C) BPS  2-1560 Bac KDM5B (PLU-1) BRIC   1-809 E. coli KDM6A (UTX) BRIC 919-1401 E. coli KDM6B (JMJD3) BPS 1043-end Bac KDM7 BRIC   1-1322 Bac(PHF8) KDM3A BPS, Bioscience,   2-end Bac (JMJD1A) US

Substrates:

BK9M3: Biotin-ARTKQTAR(KMe₃)STGGKAPRKQ-NH₂ (Caslo, Denmark)

BK9M2: Biotin-ARTKQTAR(KMe₂)STGGKAPRKQ-NH₂ (AnaSpec 64359)

BK9M1: Biotin-ARTKQTAR(KMe₁)STGGKAPRKQ-NH₂ (AnaSpec 64358)

H3K4M3B: H-ART(Kme3)QTARKSTGGKAPRKQLA-NH-Biotin (Caslo, Denmark)

BK27M3: Biotin-ATKAAR(Kme3)SAPATGGVKKPHRY-NH2? (Caslo, Denmark)

BH3K36M2: RKAAPATGGVK(Me2)KPHRYRPGTVK-(BIOTIN)? (Anaspec)

Enzyme Buffer: 50 mM Hepes (pH 7.4 or 8.0), 0.003% Tween-20, 0.1% BSA; 5μM (NH₄)₂Fe(SO₄)₂

Buffer A: 50 mM Hepes (pH 7.4 or 8.0), 0.003% Tween-20, 0.1% BSA

Substrate Solution: Substrate, 25 μM L-Asc, and 10 μM α-KG in Buffer A.

HDME Inhibition

Compound Compound Name # KDM4C KDM2B KDM5C KDM3 A KDM3B KDM4A KDM4BKDM6B PHF8 KDM6A KDM5B N-{[2-({[4- 1 + + ++ + + ++ +++ + ++ + ++(diethylamino)butyl]amino}- methyl)pyridin-4- yl]methyl}-2,2,2-trifluoroacetamide [2-({[4- 2 + + + + + + + + + + (dimethylamino)butyl]-amino}methyl)pyridin-4- yl]methanamine [2-({[3- 3 + + +++ + +++ +++ +++ + ++ (dimethylamino)propyl]- amino}methyl)pyridin-4- yl]methanamine2-({[4- 4 ++ + +++ + + ++ +++ + ++ + +++ (aminomethyl)pyridin-2-yl]methyl}amino)-N-[2- (dimethylamino)ethyl]-N- ethylacetamide [2-({[4-5 + + + + + + + + + + (diethylamino)butyl]amino}- methyl)pyridin-4-yl]methanamine N-[4- 6 + + + + + ++ +++ + ++ + + (diethylamino)butyl]-2,2,2-trifluoro-N-({4- [(trifluoroacetamido)- methyl]pyridin-2-yl}methyl)acetamide [2-({[4-(azetidin-1- 7 + + + + + +++ + + + ++yl)butyl]amino}methyl) - pyridin-4-yl]methanamine [2-({[5- 8 + ++++ + + + + + + + +++ (dimethylamino)pentyl]- amino}methyl)pyridin-4-yl]methanamine 2-({[4- 9 + + +++ + + + + + + + ++(aminomethyl)pyridin-2- yl]methyl}amino)-N-{1- [(2-methoxyphenyl)methyl]- piperidin-4-yl}acetamide N-{[2-({[4-10 + + + + + + + + + + (dimethylamino)butyl]- amino}methyl)pyridin-4-yl]methyl}cyclopropanamine N-{[2-({[3-(2- 11 + + + + + + + + + ++methylpiperidin-1- yl)propyl]amino}methyl)- pyridin-4-yl]methyl}cyclopropanamine N-({2- 12 + + + + + + +++ + + + ++[(propylamino)methyl]- pyridin-4- yl}methyl)cyclopropanamine 2-{[(4-13 + + + + + + + + + ++ {[(cyanomethyl)amino]- methyl}pyridin-2-yl)methyl]amino}-N,N- dimethylacetamide 2-{[(4-{[(2- 14 ++ + ++ + + +++ + + + ++ fluoroethyl)amino]methyl}- pyridin-2- yl)methyl]amino}-N,N-dimethylacetamide 2-({[4-({[2- 15 ++ + ++ + + + ++ + + + +++(dimethylamino)ethyl]- amino}methyl)pyridin-2- yl]methyl}amino)-N,N-dimethylacetamide {[(2S)-1- 16 + + ++ + + + + + ++ + ++benzylpyrrolidin-2- yl]methyl}[(4- {[(cyclopropylmethyl)-amino]methyl}pyridin-2- yl)methyl]amine benzyl(methyl){3-[({4- 17 ++ ++++ + + + ++ + + + +++ [(methylamino)methyl]- pyridin-2-yl}methyl)amino]propyl}- amine benzyl[3-({[4-({[2- 18 + + + + + + + + +++ (dimethylamino)ethyl]- amino}methyl)pyridin-2-yl]methyl}amino)propyl]- methylamine benzyl(3-{[(4-{[(2-19 + + + + + + + + + + ++ methoxyethyl)amino]- methyl}pyridin-2-yl)methyl]amino}propyl)- methylamine 2-[({4- 20 ++ + +++ + + +++ +++ ++++ +++ [(cyclopropylamino)- methyl]pyridin-2- yl}methyl)amino]-N-{1-[(2- methoxyphenyl)methyl]- piperidin-4-yl}acetamide2-cyclopropyl-2-({[2- 21 + + + + + + + + + + + ({[2-(dimethylamino)ethyl]- amino}methyl)pyridin-4-yl]methyl}amino)acetonitrile 2-({[2-({[3- 22 + + +++ + + + ++ + + + ++(dimethylamino)propyl]- amino}methyl)pyridin-4- yl]methyl}amino)propane-nitrile 2-[({2-[({4- 23 + + +++ + + + + + + + ++ [benzyl(cyclopropyl)-amino]butyl}amino)- methyl]pyridin-4- yl}methyl)amino]acetonitrile2-[2-({[3- 24 ++ + +++ + + ++ +++ + + + +++ (dimethylamino)propyl]-amino}methyl)pyridin-4- yl]-2- (methylamino)acetonitrile N-[(2-{[({[2-25 + + ++ + + + + + + + ++ (dimethylamino)ethyl]-(ethyl)carbamoyl}methyl)- amino]methyl}pyridin-4- yl)methyl]-2,2,2-trifluoroacetamide N-[(2-{[N-({[2- 26 + + + (dimethylamino)ethyl]-(ethyl)carbamoyl}methyl- 2,2,2- trifluoroacetamido]methyl}-pyridin-4-yl)methyl]- 2,2,2-trifluoroacetamide ({[2-({[4- 27 + + + + +++(diethylamino)butyl]- amino}methyl)pyridin-4- yl]methyl}carbamoyl)-formic acid tert-butyl ({[2-({[4- 28 + + ++ (diethylamino)butyl]-amino}methyl)pyridin-4- yl]methyl}carbamoyl) formate ethyl2-({[(2-{[({[2- 29 + + ++ (dimethylamino)ethyl](ethyl)carbamoyl}methyl)- amino]methyl}pyridin-4- yl)methyl]carbamoyl}oxy)-benzoate N-[(2-{[({[2-(azetidin-1- 30 + + + ++yl)ethyl](ethyl)carbamoyl}- methyl)amino]methyl}-pyridin-4-yl)methyl]-2,2,2- trifluoroacetamide N-[(2-{[({[2- 31 + + + ++(dimethylamino)ethyl]- (ethyl)carbamoyl}methyl)- amino]methyl}pyridin-4-yl)methyl]-2,2,3,3,4,4,4- heptafluorobutanamide N-[(2-{[({[2- 32 + + ++++ (dimethylamino)ethyl]- (ethyl)carbamoyl}methyl)-amino]methyl}pyridin-4- yl)methyl]-2,2- difluorobutanamide 2-[({4-[(N-33 +++ ++ +++ + +++ + + + + +++ cyclopropylcarboximidoyl]- pyridin-2-yl}methyl)amino]-N,N- dimethylacetamide N,N-dimethyl-2-[({4- 34 ++ ++++ + + + + + + + +++ [[(3- phenylpropyl)imino]- methyl]pyridin-2-yl}methyl)amino]acetamide N,N-dimethyl-2-[({4-[N- 35 ++ ++ +++ + + ++++++ + +++ + ++ (2- methylcyclopropyl)- carboximidoyl]pyridin-2-yl}methyl)amino]acetamide 2-[({4-[[(2- 36 ++ + +++ + + ++ +++ + ++ + +++cyclohexylethyl)imino]- methyl]pyridin-2- yl}methyl)amino]-N,N-dimethylacetamide [3- 37 + + +++ + + + ++ + + +++(dimethylamino)propyl]- ({4-[{[3- (dimethylamino)propyl]-imino}methyl]pyridin-2- yl}methyl)amine ({4-[{[2- 38 ++ ++++ + + + + + + + +++ (dimethylamino)ethyl]- imino}methyl]pyridin-2-yl}methyl)[3- (dimethylamino)propyl]- amine N-{[2-({[2- 39 + ++ +++ + +++ +++ + +++ + ++ (ethylsulfanyl)ethyl]- amino}methyl)pyridin-4-yl]methylidene}- cyclopropanamine N-{[2-({[2-(1- 40 + + +++ + + +++ + + + +++ methylpyrrolidin-2- yl)ethyl]amino}methyl)- pyridin-4-yl]methylidene}- cyclopropanamine N-({2-[({3- 41 ++ + + +++ +++ + + ++++ [benzyl(methyl)amino]- propyl}amino)methyl]- pyridin-4-yl}methylidene)- cyclopropanamine N-{[2-({[3-(pyrrolidin-1- 42 + ++++ + + + ++ + + + +++ yl)propyl]amino}methyl)- pyridin-4-yl]methylidene}- cyclopropanamine N-{[2-({[(2E)-4- 43 +++ + +++ + + +++++ + ++ + +++ (dimethylamino)but-2- en-1- yl]amino}methyl)pyridin- 4-yl]methylidene}- cyclopropanamine N-{[2-({[4-(azetidin-1- 44 +++ ++++ + + + + ++ + + +++ yl)butyl]amino}methyl)- pyridin-4-yl]methylidene}- cyclopropanamine N-{[2-({[4- 45 + + +++ + + + ++ + + +++ (dimethylamino)butyl]- amino}methyl)pyridin-4- yl]methylidene}-cyclopropanamine N-[(2-{[({4- 46 + + +++ + + ++ ++ + + + ++[(dimethylamino)methyl]- cyclohexyl}methyl)amino] - methyl}pyridin-4-yl)methylidene]- cyclopropanamine N-{[2-({[5- 47 + + +++ + + + ++ + + ++++ (dimethylamino)pentyl]- amino}methyl)pyridin-4- yl]methylidene}-cyclopropanamine 2-[({4-[N- 48 ++ + +++ + + +++ +++ + +++ + +++cyclopropylcarboximidoyl]- pyridin-2- yl}methyl)amino]-N-[4-(diethylamino)butyl]- acetamide 2-[({4-[N- 49 ++ + +++ + + +++ +++ +++ + +++ cyclopropylcarboximidoyl]- pyridin-2- yl}methyl)amino]-1-[(2R)-2-(pyrrolidin-1- ylmethyl)pyrrolidin-1- yl]ethan-1-oneN-(2-cyanoethyl)-2-[({4- 50 ++ ++ +++ + + +++ +++ + +++ + +++ [N-cyclopropylcarboximidoyl]- pyridin-2- yl}methyl)amino]-N- ethylacetamide2-[({4-[N- 51 + + +++ + + ++ +++ + ++ + +++ cyclopropylcarboximidoyl]-pyridin-2- yl}methyl)amino]-N-[(1- ethylpyrrolidin-2-yl)methyl]acetamide 2-[({4-[N- 52 ++ ++ +++ + + ++ +++ + + +++cyclopropylcarboximidoyl]- pyridin-2- yl}methyl)amino]-N-methyl-N-[3-(1H-pyrazol- 1-yl)propyl]acetamide N-(1-benzylpyrrolidin-3-53 ++ ++ +++ ++ + +++ +++ ++ +++ + +++ yl)-2-[({4-[N-cyclopropylcarboximidoyl]- pyridin-2- yl}methyl)amino]acetamide2-[({4-[N- 54 ++ + +++ + ++ ++ ++ + + + +++ cyclopropylcarboximidoyl]-pyridin-2- yl}methyl)amino]-1-(4- methylpiperazin-1- yl)ethan-1-one1-(4-benzylpipendin-1- 55 ++ + ++ + + ++ +++ + ++ + +++ yl)-2-[({4-[N-cyclopropylcarboximidoyl]- pyridin-2- yl}methyl)amino]ethan- 1-one2-[({4-[N- 56 + + ++ + + ++ ++ + ++ + ++ cyclopropylcarboximidoyl]-pyridin-2- yl}methyl)amino]-N- methyl-N-(prop-2-yn-1- yl)acetamide2-[({4-[[(2- 57 ++ ++ +++ + + ++ +++ + + + +++ cyclohexylethyl)imino]-methyl]pyridin-2- yl}methyl)amino]-N,N- diethylacetamideN,N-diethyl-2-[({4- 58 ++ ++ +++ + + + + + + + +++ [(octylimino)methyl]-pyridin-2- yl}methyl)amino]- acetamide methyl 2-[({4-[N- 59 ++ +++ + +++ +++ + ++ + +++ cyclopropylcarboximidoyl]- pyridin-2-yl}methyl)amino]acetate [4- 60 + + +++ + + +++ +++ + ++ + +++(diethylamino)butyl]({4- [[(2- methoxyethyl)imino]- methyl]pyridin-2-yl}methyl)amine 2-[{[2-({[4- 61 ++ + +++ + + + +++ + + + +++(diethylamino)butyl]amino}- methyl)pyridin-4- yl]methylidene}amino]-ethan-1-ol {[2-({[4- 62 + + +++ + + + ++ + + + +++(diethylamino)butyl]amino}- methyl)pyridin-4- yl]methylidene}(2,2,3,3,3- pentafluoropropyl)amine 2-[({4-[[(2- 63 +++ ++ +++ + + ++ + + + +++cyclohexylethyl)imino]- methyl]pyridin-2- yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N- ethylacetamide [3- 64 + + + + + ++ +++ + + ++++ (dimethylamino)propyl]- ({4- [(methoxyimino)methyl]-pyridin-2-yl}methyl)amine [4- 65 ++ + +++ + + + + + + + +++(diethylamino)butyl]({[4- (1-methylimidazolidin-2- yl)pyridin-2-yl]methy})amine N-[2- 66 +++ + +++ + + +++ +++ + ++ + +++(dimethylamino)ethyl]-N- ethyl-2-[({4-[([(2- hydroxyethyl)imino]-methyl]pyridin-2- yl}methyl)amino]acetamide (2-cyclohexylethyl)({[2- 67++ + +++ + + + + + + +++ ({[4- (diethylamino)butyl]amino} -methyl)pyridin-4- yl]methylidene})amine [4- 68 ++ + +++(diethylamino)butyl]({[4- (1-methyl-1,3-diazinan- 2-yl)pyridin-2-yl]methyl})amine N,N-diethyl-2-[({4-[{[2- 69 ++ + +++ (4-methylphenyl)ethyl]imino} - methyl]pyridin-2- yl}methyl)amino]acetamide4-[2-{[2-({[4- 70 + + ++ (diethylamino)butyl]amino}- methyl)pyridin-4-yl]methylidene}hydrazin- 1-yl]benzonitrile 3-[{[2-({[4- 71 + + +++(diethylamino)butyl]amino}- methyl)pyridin-4- yl]methylidene}amino]-propan-1-ol [4- 72 + + + ++ (diethylamino)butyl][(4- {7-oxa-9-azaspiro[4.5]decan-8- yl}pyridin-2- yl)methyl]amine 2-[{[2-({[4- 73 ++++ (diethylamino)butyl]amino}- methyl)pyridin-4- yl]methylidene}amino]-propan-1-ol 1-[{[2-({[4- 74 + +++ (diethylamino)butyl]amino}-methyl)pyridin-4- yl]methylidene}amino]- propan-2-ol 2-[{[2-({[4- 75 ++++ (diethylamino)butyl]amino}- methyl)pyridin-4- yl]methylidene}amino]-2-phenylethan-1-ol 3-[{[2-({[4- 76 + + ++ (diethylamino)butyl]amino}-methyl)pyridin-4- yl]methylidene}amino]- 2,2-dimethylpropan-1-ol(1-{[{[2-({[4- 77 + + ++ (diethylamino)butyl]amino}- methyl)pyridin-4-yl]methylidene}amino]- methyl}cyclopropyl)- methanol N-[2- 78 ++ + +++(dimethylamino)ethyl]-N- ethyl-2-[({4-[[(3- hydroxypropyl)imino]-methyl]pyridin-2- yl}methyl)amino]- acetamide N-ethyl-2-[({4-[[(2- 79++ + +++ hydroxyethyl)imino]- methyl]pyridin-2- yl}methyl)amino]-N-[(1-methylpyrrolidin-2- yl)methyl]acetamide 2-{[{[2-({[4- 80 +++ ++ + +++(diethylamino)butyl]amino}- methyl)pyridin-4- yl]methylidene}amino]-methyl}-3-phenylpropan- 1-ol 2-[({4-[[(2-cyclohexyl-3- 81 ++ + + +++hydroxypropyl)imino]- methyl]pyridin-2- yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N- ethylacetamide N-[3- 82 + + + +(dimethylamino)propyl]- N-ethyl-2-[({4-[[(2- hydroxyethyl)imino]-methyl]pyridin-2- yl}methyl)amino]acetamide N-[2- 83 ++ + +++(dimethylamino)propyl]- N-ethyl-2-[({4-[[(2- hydroxyethyl)imino]methyl]-pyridin-2- yl}methyl)amino]acetamide 1-[{[2-({[4- 84 + + + ++(diethylamino)butyl]amino}- methyl)pyridin-4- yl]methylidene}amino]-3-phenylpropan-2-ol N-{[(1S,2S)-2- 85 +++ + + +++ (dimethylamino)]-cyclopentylmethyl}- N-ethyl-2-[({4-[[(2- hydroxyethypimino]methyl]pyridin-2- yl}methyl)amino]acetamide 2-[({4-[{[3- 86 + + +++(dimethylamino)-2- hydroxypropyl]imino}- methyl]pyridin-2-yl}methyl)amino]-N-[2- (dimethylamino)ethyl]-N- ethylacetamide2-({[4-(5,5-dimethyl-1,3- 87 ++ + +++ oxazinan-2-yl)pyridin-2-yl]methyl}amino)-N-[2- (dimethylamino)ethyl]-N- ethylacetamide N-[2- 88++ + + + +++ (dimethylamino)ethyl]-N- ethyl-2-[({4-[({[1-(hydroxymethyl)- cyclopropyl]methyl}imino)- methyl]pyridin-2-yl}methyl)amino]acetamide 2-[({4-[[(2-benzyl-3- 89 ++ + + +++hydroxypropyl)imino]- methyl]pyridin-2- yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N- ethylacetamide 2-[({4-[5-benzyl-3- 90 ++ + + ++++ (trifluoroacetyl)-1,3- oxazinan-2-yl]pyridin-2-yl}methyl)amino]-N-[2- (dimethylamino)ethyl]-N- ethylacetamide N-[2- 91++ + + + +++ (dimethylamino)ethyl]-N- ethyl-2-[({4-[7-(trifluoroacetyl)-5-oxa-7- azaspiro[2.5]octan-6- yl]pyridin-2-yl}methyl)amino]acetamide N-[(2- 92 + + + + ++ fluorophenyl)methyl]-2-[({4-[[(2- hydroxyethyl)imino]- methyl]pyridin-2- yl}methyl)amino]-N-methylacetamide 2-[({2-[({2-[2- 93 + + + + + (benzyloxy)phenyl]ethyl}-amino)methyl]pyridin-4- yl}methylidene)amino]- ethan-1-olN-(2-cyanoethyl)-N- 94 ++ ++ ++ + +++ ethyl-2-[({4-[[(2-hydroxyethyl)imino]- methyl]pyridin-2- yl}methyl)amino]acetamide(2S)-2-[({4-[[(2- 95 + + + + hydroxyethyl)imino]- methyl]pyridin-2-yl}methyl)amino]-4- methyl-1-(piperidin-1- yl)pentan-1-one 2-{[({4-97 + + + + + + + + ++ [(dimethylamino)methyl]- cyclohexyl}methyl)amino]-methyl}pyridine-4- carbaldehyde 2-({[(2E)-4- 98 ++ + +++ + + + + + + ++++ (dimethylamino)but-2- en-1- yl]amino}methyl)pyridine- 4-carbaldehyde2-({[(2Z)-4- 99 + + +++ + + + + + + +++ (dimethylamino)but-2- en-1-yl]amino}methyl)pyridine- 4-carbaldehyde 2-({[(1-methylpipendin- 100 + ++++ + + + + + +++ 4- yl)methyl]amino}methyl)- pyridine-4-carbaldehydeN-[2- 101 +++ + +++ + + +++ +++ + ++ + +++ (dimethylamino)ethyl]-N-ethyl-2-{[(4- formylpyridin-2- yl)methyl]amino}acetamide2-[({2-oxo-2-[(2R)-2- 102 +++ + +++ + + + ++ + + + +++ (pyrrolidin-1-ylmethyl)pyrrolidin-1- yl]ethyl}amino)methyl]- pyridine-4-carbaldehyde2-({[2-(4- 103 + + +++ + + + + + + + ++ methylpiperazin-1-yl)-2-oxoethyl]amino}methyl)- pyridine-4-carbaldehyde N-[(1-ethylpyrrolidin-2-104 ++ + +++ + + +++ + ++ + +++ yl)methyl]-2-{[(4- formylpyridin-2-yl)methyl]amino}acetamide N,N-diethyl-2-{[(4- 105 + + + + + +++ + + ++formylpyridin-2- yl)methyl]amino}acetamide 2-({[2-(4- 106 ++ + + + + +++++ + ++ + +++ benzylpipendin-1-yl)-2- oxoethyl]amino}methyl)-pyridine-4-carbaldehyde 2-({[4- 107 + + +++ + + + +++ + + + +++(diethylamino)butyl]amino}- methyl)pyridine-4- carbaldehyde 2-({[4-108 + + +++ + + + + + + ++ (dimethylamino)butyl]-amino}methyl)pyridine-4- carbaldehyde 2-[({4- 109 ++ + + + +++ + + +++[benzyl(cyclopropyl)amino]- butyl}amino)methyl]- pyridine-4-carbaldehyde2-({[2- 110 + + +++ + + ++ + + + ++ (dimethylamino)ethyl] -amino}methyl)pyridine-4- carbaldehyde 2-({[3-(pyrrolidin-1- 111 + ++++ + + ++ + + + +++ yl)propyl]amino}methyl)- pyridine-4-carbaldehydeN-[4- 112 ++ + +++ + + +++ + +++ + (diethylamino)butyl]-2-{[(4-formylpyridin-2- yl)methyl]amino}acetamide N-(1-benzylpyrrolidin-3-113 ++ ++ + + ++ +++ + + + +++ yl)-2-{[(4-formylpyridin- 2-yl)methyl]amino}acetamide 2-({[5- 114 ++ + +++ + + + ++ + + +++(dimethylamino)pentyl]- amino}methyl)pyridine-4- carbaldehyde N-[4-115 + + +++ + + ++ + ++ + + (diethylamino)butyl]- 2,2,2-trifluoro-N-[(4-formylpyridin-2- yl)methyl]acetamide N-[2- 116 +++ + +++(diethylamino)ethyl]-N- ethyl-2-{[(4- formylpyridin-2-yl)methyl]amino}acetamide 2-[({[3-(dimethylamino)- 117 ++ + +++cyclopentyl]- methyl}amino)methyl]- pyridine-4-carbaldehydeN-[2-(dimethylamino)-2- 118 +++ + ++ + +++ methylpropyl]-N-ethyl-2-{[(4-formylpyridin-2- yl)methyl]amino}acetamide N-ethyl-2-{[(4- 119 ++++ formylpyridin-2- yl)methyl]amino}-N-[(1- methylpyrrolidin-2-yl)methyl]acetamide 2-({methyl[2-oxo-2- 120 + + ++ + + (pipendin-1-yl)ethyl]amino}methyl)- pyridine-4-carbaldehyde (a) +++: IC₅₀ < 250 nM;++: 250 nM ≦ IC₅₀ ≦ 2500 nM; +: IC₅₀ > 2500 nM

Example 3 Cell Assays for IC50 Value Determination Histone LysineDemethylase Immunofluorescence Assays for IC₅₀ Value Determination,Non-Transfected Cells

This example demonstrates the ability of compounds of the invention toinhibit demethylation of a specific histone lysine mark in a humanosteosarcoma cancer cell line.

General Method

U2OS cells were harvested and seeded into multi well plates into mediacontaining compound. The media used was DMEM containing 5% FBS andpen/strep. 20 hours after incubation of cells with compounds, the cellswere washed once in PBS, harvested by fixation with formaldehyde 4%aqueous solution, and washed in PBS. Subsequently, the cells werepermeabilized in PBS with 0.2% Triton X-100. Blocking was performed inPBS with 0.2% Triton X-100 and 5% FBS. The cells were incubated withαH3K4me3 primary antibody (Cell Signaling, #9751S) in blocking solutionover night at 4° C. After incubation with primary antibody, the cellswere washed with PBS, incubated with secondary antibody (Alexa fluor 594goat anti rabbit IgG, Invitrogen, A11012) and Hoechst, (Sigma, 33342) inblocking solution, and washed again with PBS. Finally, PBS was added andhigh throughput imaging and analysis were performed by an IN CellAnalyzer 1000 (GE Healthcare). The IC50 values were based on an averagemeasure of the staining of the H3K4me3 mark in cells.

Compound Name Compound # IC50N-{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin- 1 +++4-yl]methyl}-2,2,2-trifluoroacetamide[2-({[3-(dimethylamino)propyl]amino}methyl)pyridin- 3 +++4-yl]methanamine 2-({[4-(aminomethyl)pyridin-2-yl]methyl}amino)-N-[2- 4+++ (dimethylamino)ethyl]-N-ethylacetamideN-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-({4- 6 +++[(trifluoroacetamido)methyl]pyridin-2- yl}methyl)acetamide[2-({[4-(azetidin-1-yl)butyl]amino}methyl)pyridin-4- 7 +++yl]methanamine 2-({[4-(aminomethyl)pyridin-2-yl]methyl}amino)-N-{1- 9 ++[(2-methoxyphenyl)methyl]piperidin-4-yl]-acetamide2-{[(4-{[(cyanomethyl)amino]methyl}pyridin-2- 13 ++yl)methyl]amino}-N,N-dimethylacetamide2-[({4-[(cyclopropylamino)methyl]pyridin-2- 20 ++yl}methyl)amino]-N-{1-[(2-methoxyphenyl)methyl]piperidin-4-yl]-acetamide 2-({[2-({[3- 22 +++(dimethylamino)propyl]amino}methyl)pyridin-4-yl]methyl}amino)propanenitrile 2-[({2-[({4- 23 +++[benzyl(cyclopropyl)amino]butyl}amino)methyl]pyridin-4-yl}methyl)amino]acetonitrile 2-[2-({[3- 24 +++(dimethylamino)propyl]amino}methyl)pyridin-4-yl]-2-(methylamino)acetonitrile N-[(2-{[({[2- 25 +++(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2,2-trifluoroacetamide({[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 27 +++yl]methyl]-carbamoyl)formic acid tert-butyl({[2-({[4- 28 ++(diethylamino)butyl]amino}methyl)pyridin-4- yl]methyl}carbamoyl)formateN-[(2-{[({[2-(azetidin-1- 30 +++yl)ethyl](ethyl)carbamoyl}methyl)amino]methyl]-pyridin-4-yl)methyl]-2,2,2-trifluoroacetamide N-[(2-{[({[2- 32 +(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2-difluorobutanamide2-[({4-[(N-cyclopropylcarboximidoyl]pyridin-2- 33 ++yl}methyl)amino]-N,N-dimethylacetamide2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2- 36 +++yl}methyl)amino]-N,N-dimethylacetamide({4-[{[2-(dimethylamino)ethyl]imino]-methyl]pyridin-2- 38 +++yl}methyl)[3-(dimethylamino)propyl]amineN-{[2-({[2-(1-methylpyrrolidin-2- 40 +++yl)ethyl]amino}methyl)pyridin-4- yl]methylidene}cyclopropanamineN-{[2-({[(2E)-4-(dimethylamino)but-2-en-1- 43 +++yl]amino}methyl)pyridin-4- yl]methylidene}cyclopropanamineN-{[2-({[4-(azetidin-1-yl)butyl]amino}methyl)pyridin- 44 +++4-yl]methylidene}cyclopropanamine N-[(2-{[({4- 46 +++[(dimethylamino)methyl]cyclohexyl}methyl)amino]methyl}pyridin-4-yl)methylidene]cyclopropanamine2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2- 57 ++yl}methyl)amino]-N,N-diethylacetamideN,N-diethyl-2-[({4-[(octylimino)methyl]pyridin-2- 58 +yl}methyl)amino]acetamide2-[{[2-({[4-(diethylamino)butyl]amino]-methyl)pyridin- 61 +++4-yl]methylidene}amino]ethan-1-ol{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 62 +++yl]methylidene}(2,2,3,3,3-pentafluoropropyl)amine[3-(dimethylamino)propyl]({4- 64 ++[(methoxyimino)methyl]pyridin-2-yl}methyl)amine(2-cyclohexylethyl)({[2-({[4- 67 +++(diethylamino)butyl]amino]-methyl)pyridin-4- yl]methylidene})amine[4-(diethylamino)butyl]({[4-(1-methyl-1,3-diazinan-2- 68 +++yl)pyridin-2-yl]methyl})amine 4-[2-{[2-({[4- 70 ++(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}hydrazin-1-yl]benzonitrile1-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin- 74 +++4-yl]methylidene}amino]propan-2-ol (1-{[{[2-({[4- 77 +++(diethylamino)butyl]amino]-methyl)pyridin-4-yl]methylidene]-amino]methyl}-cyclopropyl)methanolN-ethyl-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin- 79 +++2-yl}methyl)amino]-N-[(1-methylpyrrolidin-2- yl)methyl]acetamideN-[3-(dimethylamino)propyl]-N-ethyl-2-[({4-[[(2- 82 +++hydroxyethyl)imino]methyl]pyridin-2- yl}methyl)amino]acetamideN-[2-(dimethylamino)propyl]-N-ethyl-2-[({4-[[(2- 83 +++hydroxyethyl)imino]methyl]pyridin-2- yl}methyl)amino]acetamideN-{[(1S,2S)-2-(dimethylamino)cyclopentyl]methyl}-N- 85 +++ethyl-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamideN-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[({[1- 88 +++(hydroxymethyl)cyclopropyl]methyl}imino)methyl]pyridin-2-yl}methyl)amino]acetamide2-[({4-[5-benzyl-3-(trifluoroacetyl)-1,3-oxazinan-2- 90 +++yl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamideN-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[7- 91 +++(trifluoroacetyl)-5-oxa-7-azaspiro[2.5]octan-6-yl]pyridin-2-yl}methyl)amino]acetamide 2-{[({4- 97 +++[(dimethylamino)methyl]cyclohexyl}methyl)amino]methyl}pyridine-4-carbaldehyde 2-({[(2Z)-4-(dimethylamino)but-2-en-1- 99 ++yl]amino}methyl)pyridine-4-carbaldehyde 2-({[(l-methylpiperidin-4- 100+++ yl)methyl]amino}methyl)pyridine-4-carbaldehydeN-[2-(dimethylamino)ethyl]-N-ethyl-2-{[(4- 101 +++formylpyridin-2-yl)methyl]amino}acetamide2-({[4-(diethylamino)butyl]amino}methyl)pyridine-4- 107 +++ carbaldehyde2-({[3-(pyrrolidin-1-yl)propyl]amino}methyl)pyridine-4- 111 +++carbaldehyde N-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-[(4- 115 +++formylpyridin-2-yl)methyl]acetamideN-[2-(diethylamino)ethyl]-N-ethyl-2-{[(4-formylpyridin- 116 +++2-yl)methyl]amino}acetamideN-[2-(dimethylamino)-2-methylpropyl]-N-ethyl-2-{[(4- 118 +++formylpyridin-2-yl)methyl]amino}acetamideN-ethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}-N-[(1- 119 +++methylpyrrolidin-2-yl)methyl]acetamide (a) +++: IC₅₀ <250 nM; ++: 250 nM≦ IC₅₀ ≦ 2500 nM; +: IC₅₀ >2500 nM

Example 4 Histone Lysine Demethylase Immunofluorescence Assays for IC50Value Determination

This example demonstrates the ability of the compounds of the inventionto inhibit specific histone lysine demethylases expressed in a humanosteosarcoma cell line.

General Method

U2OS cells were seeded 24 hours before transfection. Transfection wasperformed with Fugene HD transfection reagent as recommended by themanufacturer. 6 hours after transfection, the cells were harvested andseeded into multi well plates into media containing compound. The mediaused was DMEM containing 10% FBS and pen/strep. 20 hours afterincubation of cells with compounds, the cells were washed in PBS,harvested by fixation with formaldehyde 4% aqueous solution, and washedin PBS. Subsequently, the cells were permeabilized in PBS with 0.2%Triton X-100 for. Blocking was performed in PBS with 0.2% Triton X-100and 5% FBS. The cells were incubated with primary antibodies in blockingsolution over night at 4° C. The primary antibodies used in the assayswere HA.11 (Covance, MMS-101P) and the antibody detecting the markspecified in the table below. After incubation with primary antibodies,the cells were washed with PBS, incubated with secondary antibodies(Alexa fluor 594 goat anti rabbit IgG, Invitrogen, A11012; Alexa flour488 donkey anti mouse IgG, Invitrogen, A21202) and Hoechst, (Sigma,33342) in blocking solution, and washed again with PBS. Finally, PBS wasadded and high throughput imaging and analysis were performed by an INCell Analyzer 1000 (GE Healthcare). The robot software analyzedindividual cells and divided these into HA+ (transfected cells) and HA−(non-transfected cells). The IC50 values were based on an averagemeasure of the staining of the mark specified in the table below in thetransfected cells.

Primary antibody Construct Vendor/ Mark used for detec- mRNA NCBI namesource Sequence detected tion of mark ID pCMVHA Kazusa Full lengthH3K36me2 Milipore NM_012308 KDM2A 7369-I pCMVHA BRIC Full length H3K9me3Abcam NM_015061 KDM4A Ab8898 pCMVHA BRIC Full length H3K9me3 AbcamNM_014663 KDM4C Ab8898 pCMVHA BRIC Fragment H3K4me2 Millipore NM_006618KDM5B (a.a. 1-752) 07-030 pCMVHA BRIC Fragment H3K27me2 AbcamNM_001080424 KDM6B (a.a. 1026-1682) Ab24684

HDME Inhibition

Compound Compound Name # KDM4C KDM4A KDM6B KDM5B KDM2A N-{[2-({[4- 1 +++++ (diethylamino)butyl]amino}methyl) - pyridin-4-yl]methyl}-2,2,2-trifluoroacetamide [2-({[4- 2 ++ +++ (dimethylamino)butyl]amino}methyl)-pyridin-4-yl]methanamine [2-({[3- 3 + +++ (dimethylamino)propyl]amino}-methyl)pyridin-4-yl]methanamine 2-({[4-(aminomethyl)pyridin-2- 4 ++ ++++ yl]methyl}amino)-N-[2- (dimethylamino)ethyl]-N- ethylacetamide[2-({[4- 5 ++ +++ (diethylamino)butyl]amino}methyl)-pyridin-4-yl]methanamine N-[4-(diethylamino)butyl]-2,2,2- 6 + +++trifluoro-N-({4- [(trifluoroacetamido)methyl]pyridin-2-yl}methyl)acetamide [2-({[4-(azetidin-1- 7 + +++yl)butyl]amino}methyl)pyridin-4- yl]methanamine [2-({[5-(dimethylamino)-8 ++ +++ pentyl]amino}methyl) pyridin-4-yl]methanamine2-({[4-(aminomethyl)pyridin-2- 9 + + ++ yl]methyl}amino)-N-{1-[(2-methoxyphenyl)methyl]piperidin-4- yl}acetamide N-{[2-({[4- 10 + +(dimethylamino)butyl]amino}methyl)- pyridin-4-yl]methyl}cyclopropanamine N-{[2-({[3-(2-methylpiperidin-1- 11 + +yl)propyl]amino}methyl)pyridin-4- yl]methyl}cyclopropanamine N-({2-12 + + [(propylamino)methyl]pyridin-4- yl}methyl)cyclopropanamine2-{[(4-{[(cyanomethyl)amino]- 13 ++ ++ ++ methyl}pyridin-2-yl)methyl]amino}-N,N- dimethylacetamide 2-{[(4-{[(2- 14 + +fluoroethyl)amino]methyl}pyridin-2- yl)methyl]amino}-N,N-dimethylacetamide 2-({[4-({[2- 15 + + ++(dimethylamino)ethyl]amino}methyl) - pyridin-2-yl]methyl}amino)-N,N-dimethylacetamide {[(2S)-1-benzylpyrrolidin-2- 16 + yl]methyl}[(4-{[(cyclopropylmethyl)amino]methyl}- pyridin-2-yl)methyl]aminebenzyl(methyl){3-[({4- 17 + + [(methylamino)methyl]pyridin-2-yl}methyl)amino]propyl}amine benzyl(3-{[(4-{[(2- 19 + +methoxyethyl)amino]methyl}pyridin- 2-yl)methyl]amino}-propyl)methylamine 2-[({4- 20 + + ++ [(cyclopropylamino)methyl]pyridin-2-yl}methyl)amino]-N-{1-[(2- methoxyphenyl)methyl]pipendin-4-yl}acetamide 2-cyclopropyl-2-({[2-({[2- 21 + ++(dimethylamino)ethyl]amino}methyl)- pyridin-4-yl]methyl}amino)acetonitrile 2-({[2-({[3-(dimethylamino)- 22 + +++propyl]amino}methyl)- pyridin-4- yl]methyl}amino)propanenitrile2-[({2-[({4- 23 + ++ [benzyl(cyclopropyl)amino]butyl}-amino)methyl]pyridin-4- yl}methyl)amino]acetonitrile2-[2-({[3-(dimethylamino)- 24 + + +++ propyl]amino}methyl)pyridin-4-yl]-2- (methylamino)acetonitrile N-[(2-{[({[2-(dimethylamino)-25 ++ + +++ + ethyl](ethyl)carbamoyl]- methyl)amino]methyl}pyridin-4-yl)methyl]-2,2,2- trifluoroacetamide N-[(2-{[N-({[2- 26 ++ +++(dimethylamino)ethyl](ethyl)- carbamoyl]methyl)-2,2,2-trifluoroacetamido]methyl}pyridin- 4-yl)methyl]-2,2,2-trifluoroacetamide 2-[({4-[(N- 33 ++ ++cyclopropylcarboximidoyl]pyridin-2- yl}methyl)amino]-N,N-dimethylacetamide N,N-dimethyl-2-[({4-[[(3- 34 + ++phenylpropyl)imino]methyl]pyridin- 2-yl}methyl)amino]acetamideN,N-dimethyl-2-[({4-[N-(2- 35 ++ ++ methylcyclopropyl)carboximidoyl]-pyridin-2-yl}methyl)amino]acetamide 2-[({4-[[(2- 36 + + ++cyclohexylethyl)imino]methyl]pyridin- 2-yl}methyl)amino]-N,N-dimethylacetamide [3-(dimethylamino)propyl]({4-[{[3- 37 ++ +++(dimethylamino)propyl]imino}- methyl]pyridin-2-yl}methyl)amine ({4-[{[2-38 + +++ (dimethylamino)ethyl]imino}- methyl]pyridin-2-yl}methyl)[3-(dimethylamino)propyl]amine N-{[2-({[2- 39 + +(ethylsulfanyl)ethyl]amino}methyl) - pyridin-4-yl]methylidene}cyclopropanamine N-{[2-({[2-(1-methylpyrrolidin-2- 40 +++++ yl)ethyl]amino}methyl)pyridin-4- yl]methylidene}cyclopropanamineN-({2-[({3- 41 ++ + ++ [benzyl(methyl)amino]propyl}amino)-methyl]pyridin-4- yl}methylidene)cyclopropanamineN-{[2-({[3-(pyrrolidin-1- 42 ++ + + +++yl)propyl]amino}methyl)pyridin-4- yl]methylidene}cyclopropanamineN-{[2-({[(2E)-4- 43 ++ +++ (dimethylamino)but-2-en-1-yl]amino}methyl)pyridin-4- yl]methylidene}cyclopropanamineN-{[2-({[4-(dimethylamino)- 45 ++ +++ butyl]amino}methyl)- pyridin-4-yl]methylidene}cyclopropanamine N-[(2-{[({4- 46 ++ +++ +++[(dimethylamino)methyl]cyclohexyl}- methyl)amino]methyl}pyridin-4-yl)methylidene]cyclopropanamine N-{[2-({[5-(dimethylamino)- 47 ++ +++pentyl]amino}methyl)- pyridin-4- yl]methylidene}cyclopropanamine2-[({4-[N- 48 + + cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N-[4- (diethylamino)butyl]acetamide 2-[({4-[N- 49 +++++ cyclopropylcarboximidoyl]pyridin-2- yl}methyl)amino]-1-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1- yl]ethan-1-oneN-(2-cyanoethyl)-2-[({4-[N- 50 + ++ cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N-ethylacetamide 2-[({4-[N- 51 + ++cyclopropylcarboximidoyl]pyridin-2- yl}methyl)amino]-N-[(1-ethylpyrrolidin-2- yl)methyl]acetamide 2-[({4-[N- 52 + ++cyclopropylcarboximidoyl]pyridin-2- yl}methyl)amino]-N-methyl-N-[3-(1H-pyrazol-1-yl)propyl]acetamide N-(1-benzylpyrrolidin-3-yl)-2- 53 + +++ [({4-[N- cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]acetamide 2-[({4-[N- 54 + ++cyclopropylcarboximidoyl]pyridin-2- yl}methyl)amino]-1-(4-methylpiperazin-1-yl)ethan-1-one 1-(4-benzylpiperidin-1-yl)-2-[({4-55 + + [N- cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]ethan-1-one 2-[({4-[N- 56 + +cyclopropylcarboximidoyl]pyridin-2- yl}methyl)amino]-N-methyl-N-(prop-2-yn-1-yl)acetamide methyl 2-[({4-[N- 59 + +cyclopropylcarboximidoyl]pyridin-2- yl}methyl)amino]acetate[4-(diethylamino)butyl]({4-[[(2- 60 ++methoxyethyl)imino]methyl]pyridin- 2-yl}methyl)amine 2-[{[2-({[4- 61 +++++ (diethylamino)butyl]amino}methyl)- pyridin-4-yl]methylidene}amino]ethan-1-ol {[2-({[4- 62 ++ +++(diethylamino)butyl]amino}methyl)- pyridin-4- yl]methylidene}(2,2,3,3,3-pentafluoropropyl)amine 2-[({4-[[(2- 63 ++ +++cyclohexylethyl)imino]methyl]pyridin- 2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N- ethylacetamide [3-(dimethylamino)propyl]({4-64 + + [(methoxyimino)methyl]pyridin-2- yl}methyl)amine[4-(diethylamino)butyl]({[4-(1- 65 + +++methylimidazolidin-2-yl)pyridin-2- yl]methyl})amineN-[2-(dimethylamino)ethyl]-N- 66 ++ +++ ethyl-2-[({4-[([(2-hydroxyethyl)imino]methyl]pyridin- 2-yl}methyl)amino]acetamide(2-cyclohexylethyl)({[2-({[4- 67 ++ +++(diethylamino)butyl]amino}methyl)- pyridin-4-yl]methylidene})amine[4-(diethylamino)butyl]({[4-(1- 68 ++ +++methyl-1,3-diazinan-2-yl)pyridin-2- yl]methyl})amine2-[({4-[5-benzyl-3-(trifluoroacetyl)- 90 +++ + +++1,3-oxazinan-2-yl]pyridin-2- yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N- ethylacetamide N-[2-(dimethylamino)ethyl]-N- 91+++ + +++ + ethyl-2-[({4-[7-(trifluoroacetyl)-5-oxa-7-azaspiro[2.5]octan-6- yl]pyridin-2- yl}methyl)amino]acetamideN-[(2-fluorophenyl)methyl]-2-[({4- 92 + + + [[(2-hydroxyethyl)imino]methyl]pyridin- 2-yl}methyl)amino]-N- methylacetamide2-[({2-[({2-[2- 93 + + + (benzyloxy)phenyl]ethyl}amino)-methyl]pyridin-4- yl}methylidene)amino]ethan-1-olN-(2-cyanoethyl)-N-ethyl-2-[({4- 94 + ++ + [[(2-hydroxyethyl)imino]methyl]pyridin- 2-yl}methyl)amino]acetamide(2S)-2-[({4-[[(2- 95 + + + hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]-4-methyl-1- (piperidin-1-yl)pentan-1-one 2-[{4-[([(2-96 + + + hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N-methyl-N-(2- phenylethyl)acetamide 2-{[({4- 97 +++++ [(dimethylamino)methyl]cyclohexyl}- methyl)amino]methyl}pyridine-4-carbaldehyde 2-({[(2E)-4-(dimethylamino)but-2- 98 ++ +++ +++en-1-yl]amino}methyl)pyridine-4- carbaldehyde2-({[(2Z)-4-(dimethylamino)but-2- 99 + +en-1-yl]amino}methyl)pyridine-4- carbaldehyde 2-({[(1-methylpiperidin-4-100 ++ +++ yl)methyl]amino}methyl)pyridine-4- carbaldehydeN-[2-(dimethylamino)ethyl]-N- 101 ++ ++ + +++ethyl-2-{[(4-formylpyridin-2- yl)methyl]amino}acetamide2-[({2-oxo-[(2R)-2-(pyrrolidin-1- 102 ++ ++ ++ ylmethyl)pyrrolidin-1-yl]ethyl}amino)methyl]pyridine-4- carbaldehyde2-({[2-(4-methylpiperazin-1-yl)-2- 103 + ++oxoethyl]amino}methyl)pyridine-4- carbaldehydeN-[(1-ethylpyrrolidin-2-yl)methyl]- 104 + ++ 2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide N,N-diethyl-2-{[(4-formylpyridin-2- 105 + ++yl)methyl]amino}acetamide 2-({[2-(4-benzylpiperidin-1-yl)-2- 106 + +oxoethyl]amino}methyl)pyridine-4- carbaldehyde 2-({[4- 107 ++ + +++(diethylamino)butyl]amino}methyl)- pyridine-4-carbaldehyde 2-({[4- 108++ +++ (dimethylamino)butyl]amino}methyl)- pyridine-4-carbaldehyde2-[({4- 109 + + ++ [benzyl(cyclopropyl)amino]butyl} -amino)methyl]pyridine-4- carbaldehyde 2-({[2- 110 + ++(dimethylamino)ethyl]amino}methyl)- pyridine-4-carbaldehyde2-({[3-(pyrrolidin-1- 111 + + +++ yl)propyl]amino}methyl)pyridine-4-carbaldehyde N-[4-(diethylamino)butyl]-2-{[(4- 112 + + formylpyridin-2-yl)methyl]amino}acetamide N-(1-benzylpyrrolidin-3-yl)-2-{[(4- 113 + +formylpyridin-2- yl)methyl]amino}acetamide 2-({[5-(dimethylamino)- 114 ++++ pentyl]amino}methyl) pyridine-4-carbaldehydeN-[4-(diethylamino)butyl]-2,2,2- 115 ++ +++trifluoro-N-[(4-formylpyridin-2- yl)methyl]acetamideN-[2-(dimethylamino)-2- 118 ++ +++ methylpropyl]-N-ethyl-2-{[(4-formylpyridin-2- yl)methyl]amino}acetamide2-({methyl[2-oxo-2-(piperidin-1- 120 + + +yl)ethyl]amino}methyl)pyridine-4- carbaldehyde (a)+++: IC₅₀ < 250 nM;++: 250 nM ≦ IC₅₀ ≦ 2500 nM; +: IC₅₀ > 2500 nM

Example 5 Cell Proliferation Assays for EC50 Value Determination

This example demonstrates the ability of the compounds of the inventionto inhibit the proliferation of a human breast cancer cell line.

General Method

MCF7 cells were seeded in multi well plates at a density optimized togive approximately 90% confluent cells at the time of harvest. Cellswere incubated for 24 hours before addition of compound. Compounds werediluted in complete medium and added to the plates in duplicates. Thefinal concentration of DMSO was maximum 0.5%. Complete medium used wasDMEM with GlutaMAX containing 10% FBS and pen/strep.

120 hours after addition of compounds, the plates were harvested andanalyzed by ATPlite 1 Step (Perkin Elmer, cat no 6016739) according tothe manufactures recommendation.

Compound Name Compound # EC50N-{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 1 +++yl]methyl}-2,2,2-trifluoroacetamide[2-({[4-(dimethylamino)butyl]amino}methyl)pyridin-4- 2 +++yl]methanamine [2-({[3-(dimethylamino)propyl]amino}methyl)pyridin-4- 3+++ yl]methanamine 2-({[4-(aminomethyl)pyridin-2-yl]methyl}amino)-N-[2-4 +++ (dimethylamino)ethyl]-N-ethylacetamide[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 5 +++ yl]methanamineN-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-({4- 6 +++[(trifluoroacetamido)methyl]pyridin-2-yl}methyl)acetamide[2-({[4-(azetidin-1-yl)butyl]amino}methyl)pyridin-4- 7 ++ yl]methanamine[2-({[5-(dimethylamino)pentyl]amino}methyl)pyridin-4- 8 +++yl]methanamine 2-({[4-(aminomethyl)pyridin-2-yl]methyl}amino)-N-{1-[(2-9 + methoxyphenyl)methyl]piperidin-4-yl}acetamide2-({[2-({[3-(dimethylamino)propyl]amino}methyl)pyridin-4- 22 +++yl]methyl}amino)propanenitrile N-[(2-{[({[2- 25 +++(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2,2-trifluoroacetamideN-[(2-{[N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)- 26 +++2,2,2-trifluoroacetamido]methyl}pyridin-4-yl)methyl]-2,2,2-trifluoroacetamide ({[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-27 ++ yl]methyl}carbamoyl)formic acidtert-butyl({[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 28 ++yl]methyl}carbamoyl)formate ethyl 2-({[(2-{[({[2- 29 ++(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]carbamoyl}oxy)benzoate N-[(2-{[({[2-(azetidin-1- 30 +++yl)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2,2-trifluoroacetamide N-[(2-{[({[2- 31 ++(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2,3,3,4,4,4-heptafluorobutanamide N-[(2-{[({[2- 32 +(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2-difluorobutanamideN,N-dimethyl-2-[({4-[[(3-phenylpropyl)imino]methyl]pyridin-2- 34 +yl}methyl)amino]acetamide2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2- 36 ++yl}methyl)amino]-N,N-dimethylacetamide[3-(dimethylamino)propyl]({4-[{[3- 37 +++(dimethylamino)propyl]imino}methyl]pyridin-2-yl}methyl)amine({4-[{[2-(dimethylamino)ethyl]imino}methyl]pyridin-2- 38 +++yl}methyl)[3-(dimethylamino)propyl]amineN-{[2-({[2-(1-methylpyrrolidin-2-yl)ethyl]amino}methyl)pyridin- 40 ++4-yl]methylidene}cyclopropanamineN-{[2-({[3-(pyrrolidin-1-yl)propyl]amino}methyl)pyridin-4- 42 +++yl]methylidene}cyclopropanamineN-{[2-({[(2E)-4-(dimethylamino)but-2-en-1- 43 +++yl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamineN-{[2-({[4-(azetidin-1-yl)butyl]amino}methyl)pyridin-4- 44 +++yl]methylidene}cyclopropanamine N-[(2-{[({4- 46 +++[(dimethylamino)methyl]cyclohexyl}methyl)amino]methyl}pyridin-4-yl)methylidene]cyclopropanamine2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2- 57 +yl}methyl)amino]-N,N-diethylacetamideN,N-diethyl-2-[({4-[(octylimino)methyl]pyridin-2- 58 +yl}methyl)amino]acetamide [4-(diethylamino)butyl]({4-[[(2- 60 +++methoxyethyl)imino]methyl]pyridin-2-yl}methyl)amine2-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 61 +++yl]methylidene}amino]ethan-1-ol{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 62 +++yl]methylidene}(2,2,3,3,3-pentafluoropropyl)amine2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2- 63 +++yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide[3-(dimethylamino)propyl]({4-[(methoxyimino)methyl]pyridin-2- 64 +yl}methyl)amine[4-(diethylamino)butyl]({[4-(1-methylimidazolidin-2-yl)pyridin-2- 65 +++yl]methyl})amine N-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[([(2- 66 +++hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide(2-cyclohexylethyl)({[2-({[4- 67 +++(diethylamino)butyl]amino}methyl)pyridin-4- yl]methylidene})amine[4-(diethylamino)butyl]({[4-(1-methyl-1,3-diazinan-2-yl)pyridin- 68 +++2-yl]methyl})amine N,N-diethyl-2-[({4-[{[2-(4- 69 +methylphenyl)ethyl]imino}methyl]pyridin-2- yl}methyl)amino]acetamide4-[2-{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 70 ++yl]methylidene}hydrazin-1-yl]benzonitrile3-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 71 +++yl]methylidene}amino]propan-1-ol[4-(diethylamino)butyl][(4-{7-oxa-9-azaspiro[4.5]decan-8- 72 +++yl}pyridin-2-yl)methyl]amine2-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 73 +++yl]methylidene}amino]propan-1-oll-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 74 +++yl]methylidene}amino]propan-2-ol2-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 75 +++yl]methylidene}amino]-2-phenylethan-1-ol3-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 76 +++yl]methylidene}amino]-2,2-dimethylpropan-1-ol(1-{[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 77 +++yl]methylidene}amino]methyl}cyclopropyl)methanolN-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[[(3- 78 +++hydroxypropyl)imino]methyl]pyridin-2- yl}methyl)amino]acetamideN-ethyl-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2- 79 +++yl}methyl)amino]-N-[(1-methylpyrrolidin-2-yl)methyl]acetamide2-{[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 80 ++yl]methylidene}amino]methyl}-3-phenylpropan-1-ol2-[({4-[[(2-cyclohexyl-3-hydroxypropyl)imino]methyl]pyridin-2- 81 +++yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamideN-[3-(dimethylamino)propyl]-N-ethyl-2-[({4-[[(2- 82 +++hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamideN-[2-(dimethylamino)propyl]-N-ethyl-2-[({4-[[(2- 83 +++hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide1-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4- 84 +++yl]methylidene}amino]-3-phenylpropan-2-olN-{[(1S,2S)-2-(dimethylamino)cyclopentyl]methyl}-N-ethyl-2- 85 ++[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2- yl}methyl)amino]acetamide2-[({4-[{[3-(dimethylamino)-2- 86 +++hydroxypropyl]imino}methyl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide2-({[4-(5,5-dimethyl-1,3-oxazinan-2-yl)pyridin-2- 87 +++yl]methyl}amino)-N-[2-(dimethylamino)ethyl]-N-ethylacetamideN-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[({[1- 88 +++(hydroxymethyl)cyclopropyl]methyl}imino)methyl]pyridin-2-yl}methyl)amino]acetamide2-[({4-[[(2-benzyl-3-hydroxypropyl)imino]methyl]pyridin-2- 89 +++yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide2-[({4-[5-benzyl-3-(trifluoroacetyl)-1,3-oxazinan-2-yl]pyridin-2- 90 +++yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamideN-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[7-(trifluoroacetyl)-5- 91 +++oxa-7-azaspiro[2.5]octan-6-yl]pyridin-2- yl}methyl)amino]acetamideN-[(2-fluorophenyl)methyl]-2-[({4-[[(2- 92 +hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N- methylacetamide2-[({2-[({2-[2-(benzyloxy)phenyl]ethyl}amino)methyl]pyridin-4- 93 +yl}methylidene)amino]ethan-1-ol N-(2-cyanoethyl)-N-ethyl-2-[({4-[[(2-94 + hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide(2S)-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2- 95 +yl}methyl)amino]-4-methyl-1-(piperidin-1-yl)pentan-1-one 2-{[({4- 97 ++[(dimethylamino)methyl]cyclohexyl}methyl)amino]methyl}pyridine-4-carbaldehyde 2-({[(2E)-4-(dimethylamino)but-2-en-1- 98 +++yl]amino}methyl)pyridine-4-carbaldehyde2-({[(2Z)-4-(dimethylamino)but-2-en-1- 99 +yl]amino}methyl)pyridine-4-carbaldehyde2-({[(1-methylpiperidin-4-yl)methyl]amino}methyl)pyridine-4- 100 +++carbaldehyde N-[2-(dimethylamino)ethyl]-N-ethyl-2-{[(4-formylpyridin-2-101 +++ yl)methyl]amino}acetamide2-[({2-oxo-2-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1- 102 ++yl]ethyl}amino)methyl]pyridine-4-carbaldehyde2-({[4-(diethylamino)butyl]amino}methyl)pyridine-4- 107 +++ carbaldehyde2-({[3-(pyrrolidin-1-yl)propyl]amino}methyl)pyridine-4- 111 +++carbaldehydeN-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2- 115 +++yl)methyl]acetamideN-[2-(diethylamino)ethyl]-N-ethyl-2-{[(4-formylpyridin-2- 116 +++yl)methyl]amino}acetamide 2-[({[3- 117 ++(dimethylamino)cyclopentyl]methyl}amino)methyl]pyridine-4- carbaldehydeN-[2-(dimethylamino)-2-methylpropyl]-N-ethyl-2-{[(4- 118 +++formylpyridin-2-yl)methyl]amino}acetamideN-ethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}-N-[(1- 119 +++methylpyrrolidin-2-yl)methyl]acetamide (a) +++: EC₅₀ <250 nM; ++: 250 nM≦ EC₅₀ ≦ 2500 nM; +: EC₅₀ >2500 nM

Example 6 Cell Proliferation Assays for EC50 Value Determination

This example demonstrates the ability of the compounds of the inventionto inhibit the proliferation of a human cancer cell lines.

The assays were performed by the method of Example 5 by seeding therelevant cell line at a density optimized to give approximately 90%confluent cells at the time of harvest.

Compound Compound Compound Compound Compound Compound Cell Line CellType #25 #42 #61 #81 #90 #107 A375 Melanoma ++ AMO1 Plasmacytoma +++ARPE19 Retinal + + + + pigmented epithelium BT474 Mammary ++ +++ +++ductal carcinoma EJM Myeloma + + HCC1954 Breast dutal + carcinoma HEPG2Hepatocellular + + + + carcinoma JJN3 Plasma cell + leukemia JurkatAcute T cell + Clone E6-1 lymphoma K562 Chronic ++ myelogenous leukemiaKARPAS620 Plasma cell + +++ +++ +++ leukemia KMS 12 BM Myeloma +++ L1236Hodgkin's + + lymphoma L363 Plasma cell + + leukemia LP1 Myeloma + MDAMB Breast + + + 231 carcinoma MIA PACA2 Pancreas + + + epithelialcarcinoma MM1R Myeloma +++ MM1S Myeloma +++ MOLP2 Myeloma ++ + MOLP8Myeloma +++ +++ NALM6 Lymphoblastic +++ +++ leukemia NCIH929 Myeloma++ + OPM2 Myeloma ++ + ++ OVCAR-3 Ovary + RAJI Burkitt's + + + +lymphoma RPMI8226 Myeloma + + ++ + SK MM2 Plasma cell ++ leukemiaSK-MEL-28 Melanoma + SU DHL6 B cell +++ lymphoma U266 Myeloma + U2OSOsteosarcoma + + UH01 Hodgkin's + lymphoma (a) +++: EC₅₀ < 250 nM; ++:250 nM ≦ EC₅₀ ≦ 2500 nM; +: EC₅₀ > 2500 nM

Example 7 Inhibition of Tumor Growth in Mouse Xenograft Model

This example demonstrates ability of compounds of the invention toinhibit tumor growth in vivo in the OPM-2 subcutaneous mouse xenograftmodel of multiple myeloma.

Method

Briefly, NOD/SCID mice γ-irradiated with ⁶⁰Co (200 rad) (12animals/group) were inoculated subcutaneously with 8×106 OPM-2 cellsassisted with Matrigel. Dosing according to the table below started whentumors reached an average size of ˜100 mm³ (day 15). Dosing continueduntil the average size of tumors in the vehicle group reached ˜2000 mm³(day 31).

Animals were 7 weeks old female NOD/SCID mice (Mus Musculus), suppliedby Beijing HFK Bio-Technology Co. Ltd. (Beijing, china). Body weight wasapprox. 16-23 g. Before commencement of treatment, all animals wereweighed and tumor volumes were measured, and mice were assigned intogroups using randomized block design based upon their tumor volumes.

OPM-2 tumor cells were maintained in vitro in RPMI1640 mediumsupplemented with 20% fetal bovine serum at 37° C. in an atmosphere of5% CO₂ in air. The tumor cells were routinely subcultured twice weekly.The cells growing in an exponential growth phase were harvested andcounted for tumor inoculation.

Tumor sizes were measured three times weekly in two dimensions using acaliper, and the volume was expressed in mm³ using the formula: V=0.5a×b² where a and b were the long and short diameters of the tumor,respectively.

Positive control Compound Compound Compound Lenolidomide #61 #61 #61Vehicle QD × 4/ BID × 14 BID × 14 BID × 14 Days after BID × 14 week × 2i.p. 20 mg/kg i.p. 10 mg/kg i.p. 1 mg/kg inoculation Tumor Volume (mm³)15 101 ± 11 101 ± 12 101 ± 12 101 ± 11 101 ± 12 17 276 ± 19 156 ± 26 188± 25 198 ± 28 249 ± 32 19 406 ± 36 178 ± 36 292 ± 50 276 ± 44 320 ± 4521 643 ± 59 266 ± 56 440 ± 77 519 ± 67 577 ± 85 24 1047 ± 86  395 ± 92 766 ± 143 694 ± 92  951 ± 124 26 1313 ± 108  509 ± 116 1029 ± 196  928± 136 1259 ± 169 28 1776 ± 142  767 ± 169 1298 ± 236 1289 ± 178 1800 ±202 31 2473 ± 213 1148 ± 175 1573 ± 261 1996 ± 302 2864 ± 334

LIST OF REFERENCES

-   Catchpole S et al., Int. J. Oncol. 38, 1267-77, 2011-   Cloos, P. a. C. et al. (2008), Genes. Dev. 22; 115-1140-   Cloos, P. Et al., Nature 442, 307-11, 2006-   Fischle, W., et. Al., Curr. Opinion Cell Biol. 15, 172-83, 2003-   Hayami S. et al. (2010) Mol. Cancer 9-   He J et al., Blood 117 (14), 3869-80, 2011-   He J et al. Nat Struct Mol Biol 15(11), 2008-   Kelly, T. K. et al. (2010), “Epigenetic modifications as therapeutic    targets”, Nat. Biotechnol. 28; 1069-1078-   Klose, R. J. et al., Nature 442, 312-16, 2006-   Liu, G. Et al., Oncogene 28, 4491-500, 2009-   Margueron, R., et al., Curr. Opinion Genet. Dev. 15, 163-76, 2005-   Morton and Houghton, “Establishment of human tumor xenografts in    immunodeficient mice”,-   Nature Protocols, 2 (2) 247-250, 2007-   Pfau R et al., PNAS 105(6), 1907-12, 2008-   Queguiner, G. and Pastour, P., Comptes Rendus des Seances de    l'Académie des Sciences,-   Série C: Sciences Chimiques, 268(2) 182-5, 1969.-   Quina, A. S. et al. (2006), “Chromatin structure and epigenetics”,    Biochem. Pharmacol. 72; 1563-1569-   Roy et al. PerkinElmer Technical Note: AlphaLISA #12, April 2011-   Tzatsos A et al., PNAS 106 (8), 2641-6, 2009-   Yamane K. et al., Mol. Cell 25, 801-12, 2007-   Xiang Y. et al. (2007) PNAS 104

The invention claimed is:
 1. A compound of the Formula (I)

wherein Q is selected from —CH═NR¹², —W, —CH₂NHR¹³, and —CH═O; A isselected from —CH₂C(O)—, C₁₋₈ alkylene, and C₂₋₈ alkenylene, which C₁₋₈alkylene and C₂₋₈ alkenylene may optionally be substituted with one ormore R³; Y is selected from —NR⁶R⁷, —OR⁷, C₁₋₈ alkyl, C₃₋₁₀ cycloalkyl,heterocyclyl, and aryl, which C₁₋₈ alkyl, C₃₋₁₀ cycloalkyl,heterocyclyl, and aryl may optionally be substituted with one or moreR³; R¹ is selected from —H and C₁₋₈ alkyl; each R³ is independentlyselected from C₁₋₆ alkyl, C₁₋₄ hydroxyalkyl, C₃₋₁₀ cycloalkyl,—Z-heterocyclyl, —Z-aryl, NR⁶R⁷, —Z—OR⁷, halogen, and —Z—SR⁷, whereinany aryl may be substituted with one or more R⁵; each Z is independentlyselected from a single bond and C₁₋₄ alkylene; each R⁵ is independentlyselected from C₁₋₆ alkyl and C₁₋₄ alkoxy; each of R⁶ and R⁷ isindependently selected from —H, C₁₋₈ alkyl, C₁₋₄ fluoroalkyl, C₁₋₄perfluoroalkyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, and—Z-aryl, which C₁₋₈ alkyl, heterocyclyl, and aryl may optionally besubstituted with one or more independently selected R⁸; or,alternatively, R⁶ and R⁷ may together with the N-atom to which they areattached form an N-heterocyclic ring optionally substituted with one ormore independently selected R⁸; each R⁸ is independently selected fromC₁₋₆ alkyl, C₃₋₁₀ cycloalkyl, —Z-heterocyclyl, —Z-aryl, —Z—NR¹⁰R¹¹,halogen, and —CN, which C₃₋₁₀ cycloalkyl, heterocyclyl, and aryl mayoptionally be substituted with one or more selected from C₁₋₄ alkyl,—Z—NR¹⁰R¹¹, —Z—OR⁹, halogen, and —Z—SOR⁹; each R⁹ is independentlyselected C₁₋₈ alkyl; and each of R¹⁰ and R¹¹ is independently selectedC₁₋₆ alkyl, or, alternatively, R¹⁰ and R¹¹ may together with the N-atomto which they are attached form an N-heterocyclic ring; with the provisothat Y is not H when A is —CH₂—; when Q is —CH═NR¹², R¹² is selectedfrom C₁₋₁₀ alkyl, C₃₋₁₀ cycloalkyl, —Z-aryl, —Z—NR⁶R⁷, and —Z—OR⁷, whichC₁₋₁₀ alkyl, C₃₋₁₀ cycloalkyl, and aryl may optionally be substitutedwith one or more R³; when Q is —CH₂NHR¹³, R¹³ is selected from hydrogen,—C(O)R⁷, —C(O)C(O)OR⁷, C₁₋₈ alkyl, and C₃₋₁₀ cycloalkyl, which C₁₋₈alkyl may optionally be substituted with one or more independentlyselected R⁸, with the proviso that C₁₋₈ alkyl is not substituted with—Z-aryl; when Q is W, W is selected from an 1,3-diaza-C₅₋₇-cycloalk-2-ylgroup which is N-substituted with R¹⁶ and optionally further substitutedwith one or more R³; a 1,3-thiaza-C₅₋₇-cycloalk-2-yl group which isN-substituted with R¹⁶ and optionally further substituted with one ormore R³; and an 1,3-oxaza-C₅₋₇-cycloalk-2-yl group which isN-substituted with R¹⁶ and optionally further substituted with one ormore R³ wherein in all three instances two R³'s on the same carbon atommay together form a spiro group; R¹⁶ is selected from hydrogen and—C(O)R⁷; and each aryl is phenyl and each heterocyclyl comprises 1 or 2nitrogen atoms and 4 or 5 total ring atoms; or an isomer or a mixture ofisomers thereof, or a pharmaceutically acceptable salt, or solvatethereof.
 2. A compound according to claim 1, wherein A is selected from—CHR²C(O)— and C₁₋₈ alkylene.
 3. A compound according to claim 1,wherein Y is —NR⁶R⁷.
 4. A compound according to claim 3, wherein A is—CHR²C(O)—.
 5. A compound according to claim 4, wherein A is —CH₂C(O)—.6. A compound according to claim 2, wherein Y is

wherein n is from 1 to 3 and each of R⁶, R¹⁰ and R¹¹ independently is asdefined in claim
 1. 7. A compound according to claim 6, wherein Y is

wherein n is from 1 to 3 and each of R¹⁰ and R¹¹ independently is asdefined in claim
 1. 8. A compound according to claim 6, wherein Y is

wherein n is from 1 to 3 and each m independently is from 0 to
 2. 9. Acompound according to claim 1, wherein Y is selected from heterocyclyland aryl, each of which may be optionally substituted with one or moreR³.
 10. A compound according to claim 1, wherein R¹³ is H.
 11. Acompound according to claim 1, wherein Q is of the formula

wherein R¹⁸ and R¹⁹ are hydrogen, or together form a1,3-diaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³; a1,3-thiaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³; an1,3-oxaza-C₅₋₇-cycloalk-2-yl group which is N-substituted with R¹⁶ andoptionally further substituted with one or more R³, wherein in all threeinstances two R³'s on the same carbon atom may together form a spirogroup.
 12. A compound selected fromN-{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methyl}-2,2,2-trifluoroacetamide;[2-({[4-(dimethylamino)butyl]amino}methyl)pyridin-4-yl]methanamine;[2-({[3-(dimethylamino)propyl]amino}methyl)pyridin-4-yl]methanamine;2-({[4-(aminomethyl)pyridin-2-yl]methyl}amino)-N-[2-(dimethylamino)ethyl]-N-ethylacetamide;[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methanamine;N-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-({4-[(trifluoroacetamido)methyl]pyridin-2-yl}methyl)acetamide;[2-({[4-(azetidin-1-yl)butyl]amino}methyl)pyridin-4-yl]methanamine;[2-({[5-(dimethylamino)pentyl]amino}methyl)pyridin-4-yl]methanamine;2-({[4-(aminomethyl)pyridin-2-yl]methyl}amino)-N-{1-[(2-methoxyphenyl)methyl]piperidin-4-yl}acetamide;N-{[2-({[4-(dimethylamino)butyl]amino}methyl)pyridin-4-yl]methyl}cyclopropanamine;N-{[2-({[3-(2-methylpiperidin-1-yl)propyl]amino}methyl)pyridin-4-yl]methyl}cyclopropanamine;N-({2-[(propylamino)methyl]pyridin-4-yl}methyl)cyclopropanamine;2-{[(4-{[(cyanomethyl)amino]methyl}pyridin-2-yl)methyl]amino}-N,N-dimethylacetamide;2-{[(4-{[(2-fluoroethyl)amino]methyl}pyridin-2-yl)methyl]amino}-N,N-dimethylacetamide;2-({[4-({[2-(dimethylamino)ethyl]amino}methyl)pyridin-2-yl]methyl}amino)-N,N-dimethylacetamide;{[(2S)-1-benzylpyrrolidin-2-yl]methyl}[(4-{[(cyclopropylmethyl)amino]methyl}pyridin-2-yl)methyl]amine;benzyl(methyl){3-[({4-[(methylamino)methyl]pyridin-2-yl}methyl)amino]propyl}amine;benzyl[3-({[4-({[2-(dimethylamino)ethyl]amino}methyl)pyridin-2-yl]methyl}amino)propyl]methylamine;benzyl(3-{[(4-{[(2-methoxyethyl)amino]methyl}pyridin-2-yl)methyl]amino}propyl)methylamine;2-[({4-[(cyclopropylamino)methyl]pyridin-2-yl}methyl)amino]-N-{1-[(2-methoxyphenyl)methyl]piperidin-4-yl}acetamide;2-cyclopropyl-2-({[2-({[2-(dimethylamino)ethyl]amino}methyl)pyridin-4-yl]methyl}amino)acetonitrile;2-({[2-({[3-(dimethylamino)propyl]amino}methyl)pyridin-4-yl]methyl}amino)propanenitrile;2-[({2-[({4-[benzyl(cyclopropyl)amino]butyl}amino)methyl]pyridin-4-yl}methyl)amino]acetonitrile;2-[2-({[3-(dimethylamino)propyl]amino}methyl)pyridin-4-yl]-2-(methylamino)acetonitrile;N-[(2-{[({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2,2-trifluoroacetamide;N-[(2-{[N-({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)-2,2,2-trifluoroacetamido]methyl}pyridin-4-yl)methyl]-2,2,2-trifluoroacetamide;({[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methyl}carbamoyl)formicacid; tert-butyl({[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methyl}carbamoyl)formate;ethyl2-({[(2-{[({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]carbamoyl}oxy)benzoate;N-[(2-{[({[2-(azetidin-1-yl)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2,2-trifluoroacetamide;N-[(2-{[({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2,3,3,4,4,4-heptafluorobutanamide;N-[(2-{[({[2-(dimethylamino)ethyl](ethyl)carbamoyl}methyl)amino]methyl}pyridin-4-yl)methyl]-2,2-difluorobutanamide;2-[({4-[(N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N,N-dimethylacetamide;N,N-dimethyl-2-[({4-[[(3-phenylpropyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide;N,N-dimethyl-2-[({4-[N-(2-methylcyclopropyl)carboximidoyl]pyridin-2-yl}methyl)amino]acetamide;2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N,N-dimethylacetamide;[3-(dimethylamino)propyl]({4-[{[3-(dimethylamino)propyl]imino}methyl]pyridin-2-yl}methyl)amine;({4-[{[2-(dimethylamino)ethyl]imino}methyl]pyridin-2-yl}methyl)[3-(dimethylamino)propyl]amine;N-{[2-({[2-(ethylsulfanyl)ethyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine;N-{[2-({[2-(1-methylpyrrolidin-2-yl)ethyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine;N-({2-[({3-[benzyl(methyl)amino]propyl}amino)methyl]pyridin-4-yl}methylidene)cyclopropanamine;N-{[2-({[3-(pyrrolidin-1-yl)propyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine;N-{[2-({[(2E)-4-(dimethylamino)but-2-en-1-yl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine;N-{[2-({[4-(azetidin-1-yl)butyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine;N-{[2-({[4-(dimethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine;N-[(2-{[({4-[(dimethylamino)methyl]cyclohexyl}methyl)amino]methyl}pyridin-4-yl)methylidene]cyclopropanamine;N-{[2-({[5-(dimethylamino)pentyl]amino}methyl)pyridin-4-yl]methylidene}cyclopropanamine;2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N-[4-(diethylamino)butyl]acetamide;2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-1-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethan-1-one;N-(2-cyanoethyl)-2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N-ethylacetamide;2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N-[(1-ethylpyrrolidin-2-yl)methyl]acetamide;2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N-methyl-N-[3-(1H-pyrazol-1-yl)propyl]acetamide;N-(1-benzylpyrrolidin-3-yl)-2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]acetamide;2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-1-(4-methylpiperazin-1-yl)ethan-1-one;1-(4-benzylpiperidin-1-yl)-2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]ethan-1-one;2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]-N-methyl-N-(prop-2-yn-1-yl)acetamide;2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N,N-diethylacetamide;N,N-diethyl-2-[({4-[(octylimino)methyl]pyridin-2-yl}methyl)amino]acetamidemethyl2-[({4-[N-cyclopropylcarboximidoyl]pyridin-2-yl}methyl)amino]acetate;[4-(diethylamino)butyl]({4-[[(2-methoxyethyl)imino]methyl]pyridin-2-yl}methyl)amine;2-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]ethan-1-ol;{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}(2,2,3,3,3-pentafluoropropyl)amine;2-[({4-[[(2-cyclohexylethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide;[3-(dimethylamino)propyl]({4-[(methoxyimino)methyl]pyridin-2-yl}methyl)amine;[4-(diethylamino)butyl]({[4-(1-methylimidazolidin-2-yl)pyridin-2-yl]methyl})amine;N-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[([(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide;(2-cyclohexylethyl)({[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene})amine;[4-(diethylamino)butyl]({[4-(1-methyl-1,3-diazinan-2-yl)pyridin-2-yl]methyl})amine;N,N-diethyl-2-[({4-[{[2-(4-methylphenyl)ethyl]imino}methyl]pyridin-2-yl}methyl)amino]acetamide;4-[2-{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}hydrazin-1-yl]benzonitrile;3-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]propan-1-ol;[4-(diethylamino)butyl][(4-{7-oxa-9-azaspiro[4.5]decan-8-yl}pyridin-2-yl)methyl]amine;2-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]propan-1-ol;1-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]propan-2-ol;2-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]-2-phenylethan-1-ol;3-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]-2,2-dimethylpropan-1-ol;(1-{[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]methyl}cyclopropyl)methanol;N-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[[(3-hydroxypropyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide;N-ethyl-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N-[(1-methylpyrrolidin-2-yl)methyl]acetamide;2-{[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]methyl}-3-phenylpropan-1-ol;2-[({4-[[(2-cyclohexyl-3-hydroxypropyl)imino]methyl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide;N-[3-(dimethylamino)propyl]-N-ethyl-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide;N-[2-(dimethylamino)propyl]-N-ethyl-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide;1-[{[2-({[4-(diethylamino)butyl]amino}methyl)pyridin-4-yl]methylidene}amino]-3-phenylpropan-2-ol;N-{[(1S,2S)-2-(dimethylamino)cyclopentyl]methyl}-N-ethyl-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide;2-[({4-[{[3-(dimethylamino)-2-hydroxypropyl]imino}methyl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide;2-({[4-(5,5-dimethyl-1,3-oxazinan-2-yl)pyridin-2-yl]methyl}amino)-N-[2-(dimethylamino)ethyl]-N-ethylacetamide;N-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[({[1-(hydroxymethyl)cyclopropyl]methyl}imino)methyl]pyridin-2-yl}methyl)amino]acetamide;2-[({4-[[(2-benzyl-3-hydroxypropyl)imino]methyl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide;2-[({4-[5-benzyl-3-(trifluoroacetyl)-1,3-oxazinan-2-yl]pyridin-2-yl}methyl)amino]-N-[2-(dimethylamino)ethyl]-N-ethylacetamide;N-[2-(dimethylamino)ethyl]-N-ethyl-2-[({4-[7-(trifluoroacetyl)-5-oxa-7-azaspiro[2.5]octan-6-yl]pyridin-2-yl}methyl)amino]acetamide;N-[(2-fluorophenyl)methyl]-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N-methylacetamide;2-[({2-[({2-[2-(benzyloxy)phenyl]ethyl}amino)methyl]pyridin-4-yl}methylidene)amino]ethan-1-ol;N-(2-cyanoethyl)-N-ethyl-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]acetamide;(2S)-2-[({4-[[(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]-4-methyl-1-(piperidin-1-yl)pentan-1-one;2-[{4-[([(2-hydroxyethyl)imino]methyl]pyridin-2-yl}methyl)amino]-N-methyl-N-(2-phenylethyl)acetamide;2-{[({4-[(dimethylamino)methyl]cyclohexyl}methyl)amino]methyl}pyridine-4-carbaldehyde;2-({[(2E)-4-(dimethylamino)but-2-en-1-yl]amino}methyl)pyridine-4-carbaldehyde;2-({[(2Z)-4-(dimethylamino)but-2-en-1-yl]amino}methyl)pyridine-4-carbaldehyde;2-({[(1-methylpiperidin-4-yl)methyl]amino}methyl)pyridine-4-carbaldehyde;N-[2-(dimethylamino)ethyl]-N-ethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide;2-[({2-oxo-2-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl]ethyl}amino)methyl]pyridine-4-carbaldehyde;2-({[2-(4-methylpiperazin-1-yl)-2-oxoethyl]amino}methyl)pyridine-4-carbaldehyde;N-[(1-ethylpyrrolidin-2-yl)methyl]-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide;N,N-diethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide;2-({[2-(4-benzylpiperidin-1-yl)-2-oxoethyl]amino}methyl)pyridine-4-carbaldehyde;2-({[4-(diethylamino)butyl]amino}methyl)pyridine-4-carbaldehyde;2-({[4-(dimethylamino)butyl]amino}methyl)pyridine-4-carbaldehyde;2-[({4-[benzyl(cyclopropyl)amino]butyl}amino)methyl]pyridine-4-carbaldehyde;2-({[2-(dimethylamino)ethyl]amino}methyl)pyridine-4-carbaldehyde;2-({[3-(pyrrolidin-1-yl)propyl]amino}methyl)pyridine-4-carbaldehyde;N-[4-(diethylamino)butyl]-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide;N-(1-benzylpyrrolidin-3-yl)-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide;2-({[5-(dimethylamino)pentyl]amino}methyl)pyridine-4-carbaldehyde;N-[4-(diethylamino)butyl]-2,2,2-trifluoro-N-[(4-formylpyridin-2-yl)methyl]acetamide;N-[2-(diethylamino)ethyl]-N-ethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide;2-[({[3-(dimethylamino)cyclopentyl]methyl}amino)methyl]pyridine-4-carbaldehyde;N-[2-(dimethylamino)-2-methylpropyl]-N-ethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}acetamide;N-ethyl-2-{[(4-formylpyridin-2-yl)methyl]amino}-N-[(1-methylpyrrolidin-2-yl)methyl]acetamide;2-({methyl[2-oxo-2-(piperidin-1-yl)ethyl]amino}methyl)pyridine-4-carbaldehyde;and pharmaceutically acceptable salts thereof.
 13. A compound accordingto claim 12, in the form of an oxalate salt.
 14. A compound according toclaim 12, in the form of a citrate salt.
 15. A compound according toclaim 12, in the form of a fumarate salt.
 16. A compound according toclaim 12, in the form of an ascorbate salt.
 17. A pharmaceuticalcomposition comprising at least one compound of claim 1 and one or morepharmaceutically acceptable excipients, diluents or carriers.
 18. Apharmaceutical composition according to claim 17, which comprises one ormore further active substances.
 19. A method of treating cancer in asubject, comprising administering to said subject a therapeuticallyeffective amount of at least one compound of claim 1, wherein the canceris squamous cell carcinoma.
 20. The method of claim 19, wherein thesquamous cell carcinoma is located in the skin, lips, mouth, esophagus,urinary bladder, prostate, lungs, vagina, or cervix.
 21. A method oftreating cancer in a subject, comprising administering to said subject atherapeutically effective amount of at least one compound of claim 12,wherein the cancer is squamous cell carcinoma.
 22. The method of claim21, wherein the squamous cell carcinoma is located in the skin, lips,mouth, esophagus, urinary bladder, prostate, lungs, vagina, or cervix.23. A method of treating cancer in a subject, comprising administeringto said subject a therapeutically effective amount of at least onecompound of claim 1, wherein the cancer is selected from the groupconsisting of leukemia, lymphoma, oral cancer, laryngeal cancer,esophageal cancer, prostate cancer, bladder cancer, renal cancer,uterine cancer, ovarian cancer, testicular cancer, rectal cancer, coloncancer, lung cancer, brain cancer, breast cancer, pancreatic cancer,stomach cancer, liver cancer, thyroid cancer, melanoma, and multiplemyeloma.
 24. A method of treating cancer in a subject, comprisingadministering to said subject a therapeutically effective amount of atleast one compound of claim 12, wherein the cancer is selected from thegroup consisting of leukemia, lymphoma, oral cancer, laryngeal cancer,esophageal cancer, prostate cancer, bladder cancer, renal cancer,uterine cancer, ovarian cancer, testicular cancer, rectal cancer, coloncancer, lung cancer, brain cancer, breast cancer, pancreatic cancer,stomach cancer, liver cancer, thyroid cancer, melanoma, and multiplemyeloma.
 25. A method for inhibiting a histone demethylase, comprisingcontacting a cell with a compound of claim 1 in an amount effective toproduce a concentration sufficient to inhibit the demethylation of ahistone in a cell.
 26. The method of claim 25, wherein the histonedemethylase is a member of the KDM6 family, KDM5 family, KDM4 family,KDM3 family, or KDM2 family.
 27. A method for inhibiting a histonedemethylase, comprising contacting a cell with a compound of claim 12 inan amount effective to produce a concentration sufficient to inhibit thedemethylation of a histone in a cell.
 28. The method of claim 27,wherein the histone demethylase is a member of the KDM6 family, KDM5family, KDM4 family, KDM3 family, or KDM2 family.